PMC:1382200 JSON TXT 7 Projects

Annnotations TAB TSV DIC JSON TextAE

Id	Subject	Object	Predicate	Lexical cue
T665	0-1	NN	denotes	m
T667	0-71	sentence	denotes	m-Calpain is required for preimplantation embryonic development in mice
T668	1-2	HYPH	denotes	-
T666	2-9	NN	denotes	Calpain
T670	10-12	VBZ	denotes	is
T669	13-21	VBN	denotes	required
T671	22-25	IN	denotes	for
T672	26-41	NN	denotes	preimplantation
T674	42-51	JJ	denotes	embryonic
T673	52-63	NN	denotes	development
T675	64-66	IN	denotes	in
T676	67-71	NNS	denotes	mice
T677	71-72	sentence	denotes
T678	82-92	sentence	denotes	Background
T679	82-92	NN	denotes	Background
T680	92-250	sentence	denotes	μ-calpain and m-calpain are ubiquitously expressed proteases implicated in cellular migration, cell cycle progression, degenerative processes and cell death.
T681	93-94	NN	denotes	μ
T683	94-95	HYPH	denotes	-
T682	95-102	NN	denotes	calpain
T685	103-106	CC	denotes	and
T686	107-108	NN	denotes	m
T688	108-109	HYPH	denotes	-
T687	109-116	NN	denotes	calpain
T684	117-120	VBP	denotes	are
T689	121-133	RB	denotes	ubiquitously
T690	134-143	VBN	denotes	expressed
T691	144-153	NNS	denotes	proteases
T692	154-164	VBN	denotes	implicated
T693	165-167	IN	denotes	in
T694	168-176	JJ	denotes	cellular
T695	177-186	NN	denotes	migration
T696	186-188	,	denotes	,
T697	188-192	NN	denotes	cell
T698	193-198	NN	denotes	cycle
T699	199-210	NN	denotes	progression
T700	210-212	,	denotes	,
T701	212-224	JJ	denotes	degenerative
T702	225-234	NNS	denotes	processes
T703	235-238	CC	denotes	and
T704	239-243	NN	denotes	cell
T705	244-249	NN	denotes	death
T706	249-250	.	denotes	.
T707	250-424	sentence	denotes	These heterodimeric enzymes are composed of distinct catalytic subunits, encoded by Capn1 (μ-calpain) or Capn2 (m-calpain), and a common regulatory subunit encoded by Capn4.
T708	251-256	DT	denotes	These
T710	257-270	JJ	denotes	heterodimeric
T709	271-278	NNS	denotes	enzymes
T712	279-282	VBP	denotes	are
T711	283-291	VBN	denotes	composed
T713	292-294	IN	denotes	of
T714	295-303	JJ	denotes	distinct
T716	304-313	JJ	denotes	catalytic
T715	314-322	NNS	denotes	subunits
T717	322-324	,	denotes	,
T718	324-331	VBN	denotes	encoded
T719	332-334	IN	denotes	by
T720	335-340	NN	denotes	Capn1
T721	341-342	-LRB-	denotes	(
T722	342-343	NN	denotes	μ
T724	343-344	HYPH	denotes	-
T723	344-351	NN	denotes	calpain
T725	351-352	-RRB-	denotes	)
T726	353-355	CC	denotes	or
T727	356-361	NN	denotes	Capn2
T728	362-363	-LRB-	denotes	(
T729	363-364	NN	denotes	m
T731	364-365	HYPH	denotes	-
T730	365-372	NN	denotes	calpain
T732	372-373	-RRB-	denotes	)
T733	373-375	,	denotes	,
T734	375-378	CC	denotes	and
T735	379-380	DT	denotes	a
T737	381-387	JJ	denotes	common
T738	388-398	JJ	denotes	regulatory
T736	399-406	NN	denotes	subunit
T739	407-414	VBN	denotes	encoded
T740	415-417	IN	denotes	by
T741	418-423	NN	denotes	Capn4
T742	423-424	.	denotes	.
T743	424-645	sentence	denotes	Disruption of the mouse Capn4 gene abolished both μ-calpain and m-calpain activity, and resulted in embryonic lethality, thereby suggesting essential roles for one or both of these enzymes during mammalian embryogenesis.
T744	425-435	NN	denotes	Disruption
T746	436-438	IN	denotes	of
T747	439-442	DT	denotes	the
T749	443-448	NN	denotes	mouse
T750	449-454	NN	denotes	Capn4
T748	455-459	NN	denotes	gene
T745	460-469	VBD	denotes	abolished
T751	470-474	CC	denotes	both
T753	475-476	NN	denotes	μ
T754	476-477	HYPH	denotes	-
T752	477-484	NN	denotes	calpain
T756	485-488	CC	denotes	and
T757	489-490	NN	denotes	m
T759	490-491	HYPH	denotes	-
T758	491-498	NN	denotes	calpain
T755	499-507	NN	denotes	activity
T760	507-509	,	denotes	,
T761	509-512	CC	denotes	and
T762	513-521	VBD	denotes	resulted
T763	522-524	IN	denotes	in
T764	525-534	JJ	denotes	embryonic
T765	535-544	NN	denotes	lethality
T766	544-546	,	denotes	,
T767	546-553	RB	denotes	thereby
T768	554-564	VBG	denotes	suggesting
T769	565-574	JJ	denotes	essential
T770	575-580	NNS	denotes	roles
T771	581-584	IN	denotes	for
T772	585-588	CD	denotes	one
T773	589-591	CC	denotes	or
T774	592-596	DT	denotes	both
T775	597-599	IN	denotes	of
T776	600-605	DT	denotes	these
T777	606-613	NNS	denotes	enzymes
T778	614-620	IN	denotes	during
T779	621-630	JJ	denotes	mammalian
T780	631-644	NN	denotes	embryogenesis
T781	644-645	.	denotes	.
T782	645-853	sentence	denotes	Disruption of the Capn1 gene produced viable, fertile mice implying that either m-calpain could compensate for the loss of μ-calpain, or that the loss of m-calpain was responsible for death of Capn4-/- mice.
T783	646-656	NN	denotes	Disruption
T785	657-659	IN	denotes	of
T786	660-663	DT	denotes	the
T788	664-669	NN	denotes	Capn1
T787	670-674	NN	denotes	gene
T784	675-683	VBD	denotes	produced
T789	684-690	JJ	denotes	viable
T791	690-692	,	denotes	,
T792	692-699	JJ	denotes	fertile
T790	700-704	NNS	denotes	mice
T793	705-713	VBG	denotes	implying
T794	714-718	IN	denotes	that
T796	719-725	DT	denotes	either
T797	726-727	NN	denotes	m
T799	727-728	HYPH	denotes	-
T798	728-735	NN	denotes	calpain
T800	736-741	MD	denotes	could
T795	742-752	VB	denotes	compensate
T801	753-756	IN	denotes	for
T802	757-760	DT	denotes	the
T803	761-765	NN	denotes	loss
T804	766-768	IN	denotes	of
T805	769-770	NN	denotes	μ
T807	770-771	HYPH	denotes	-
T806	771-778	NN	denotes	calpain
T808	778-780	,	denotes	,
T809	780-782	CC	denotes	or
T810	783-787	IN	denotes	that
T812	788-791	DT	denotes	the
T813	792-796	NN	denotes	loss
T814	797-799	IN	denotes	of
T815	800-801	NN	denotes	m
T817	801-802	HYPH	denotes	-
T816	802-809	NN	denotes	calpain
T811	810-813	VBD	denotes	was
T818	814-825	JJ	denotes	responsible
T819	826-829	IN	denotes	for
T820	830-835	NN	denotes	death
T821	836-838	IN	denotes	of
T822	839-844	NN	denotes	Capn4
T824	844-845	SYM	denotes	-
T825	845-846	HYPH	denotes	/
T826	846-847	SYM	denotes	-
T823	848-852	NNS	denotes	mice
T827	852-853	.	denotes	.
T828	853-862	sentence	denotes	Results
T829	855-862	NNS	denotes	Results
T830	862-1069	sentence	denotes	To distinguish between the alternatives described above, we deleted an essential coding region in the mouse Capn2 gene in embryonic stems cells and transmitted this mutant allele through the mouse germline.
T831	863-865	TO	denotes	To
T832	866-877	VB	denotes	distinguish
T834	878-885	IN	denotes	between
T835	886-889	DT	denotes	the
T836	890-902	NNS	denotes	alternatives
T837	903-912	VBN	denotes	described
T838	913-918	RB	denotes	above
T839	918-920	,	denotes	,
T840	920-922	PRP	denotes	we
T833	923-930	VBD	denotes	deleted
T841	931-933	DT	denotes	an
T843	934-943	JJ	denotes	essential
T844	944-950	NN	denotes	coding
T842	951-957	NN	denotes	region
T845	958-960	IN	denotes	in
T846	961-964	DT	denotes	the
T848	965-970	NN	denotes	mouse
T849	971-976	NN	denotes	Capn2
T847	977-981	NN	denotes	gene
T850	982-984	IN	denotes	in
T851	985-994	JJ	denotes	embryonic
T853	995-1000	VBZ	denotes	stems
T852	1001-1006	NNS	denotes	cells
T854	1007-1010	CC	denotes	and
T855	1011-1022	VBN	denotes	transmitted
T856	1023-1027	DT	denotes	this
T858	1028-1034	NN	denotes	mutant
T857	1035-1041	NN	denotes	allele
T859	1042-1049	IN	denotes	through
T860	1050-1053	DT	denotes	the
T862	1054-1059	NN	denotes	mouse
T861	1060-1068	NN	denotes	germline
T863	1068-1069	.	denotes	.
T864	1069-1175	sentence	denotes	Breeding of heterozygous animals failed to produce homozygous mutant live offspring or implanted embryos.
T865	1070-1078	NN	denotes	Breeding
T867	1079-1081	IN	denotes	of
T868	1082-1094	JJ	denotes	heterozygous
T869	1095-1102	NNS	denotes	animals
T866	1103-1109	VBD	denotes	failed
T870	1110-1112	TO	denotes	to
T871	1113-1120	VB	denotes	produce
T872	1121-1131	JJ	denotes	homozygous
T874	1132-1138	NN	denotes	mutant
T875	1139-1143	JJ	denotes	live
T873	1144-1153	NN	denotes	offspring
T876	1154-1156	CC	denotes	or
T877	1157-1166	VBN	denotes	implanted
T878	1167-1174	NNS	denotes	embryos
T879	1174-1175	.	denotes	.
T880	1175-1332	sentence	denotes	A nested PCR genotyping protocol was established, and homozygous preimplantation mutant embryos were detected at the morula but not at the blastocyts stage.
T881	1176-1177	DT	denotes	A
T883	1178-1184	VBN	denotes	nested
T884	1185-1188	NN	denotes	PCR
T885	1189-1199	NN	denotes	genotyping
T882	1200-1208	NN	denotes	protocol
T887	1209-1212	VBD	denotes	was
T886	1213-1224	VBN	denotes	established
T888	1224-1226	,	denotes	,
T889	1226-1229	CC	denotes	and
T890	1230-1240	JJ	denotes	homozygous
T892	1241-1256	NN	denotes	preimplantation
T893	1257-1263	NN	denotes	mutant
T891	1264-1271	NNS	denotes	embryos
T895	1272-1276	VBD	denotes	were
T894	1277-1285	VBN	denotes	detected
T896	1286-1288	IN	denotes	at
T897	1289-1292	DT	denotes	the
T898	1293-1299	NN	denotes	morula
T899	1300-1303	CC	denotes	but
T900	1304-1307	RB	denotes	not
T901	1308-1310	IN	denotes	at
T902	1311-1314	DT	denotes	the
T903	1315-1325	NNS	denotes	blastocyts
T904	1326-1331	NN	denotes	stage
T905	1331-1332	.	denotes	.
T906	1332-1344	sentence	denotes	Conclusion
T907	1334-1344	NN	denotes	Conclusion
T908	1344-1490	sentence	denotes	We conclude that homozygous disruption of the Capn2 gene results in pre-implantation embryonic lethality between the morula and blastocyst stage.
T909	1345-1347	PRP	denotes	We
T910	1348-1356	VBP	denotes	conclude
T911	1357-1361	IN	denotes	that
T913	1362-1372	JJ	denotes	homozygous
T914	1373-1383	NN	denotes	disruption
T915	1384-1386	IN	denotes	of
T916	1387-1390	DT	denotes	the
T918	1391-1396	NN	denotes	Capn2
T917	1397-1401	NN	denotes	gene
T912	1402-1409	VBZ	denotes	results
T919	1410-1412	IN	denotes	in
T920	1413-1429	JJ	denotes	pre-implantation
T922	1430-1439	JJ	denotes	embryonic
T921	1440-1449	NN	denotes	lethality
T923	1450-1457	IN	denotes	between
T924	1458-1461	DT	denotes	the
T926	1462-1468	NN	denotes	morula
T927	1469-1472	CC	denotes	and
T928	1473-1483	NN	denotes	blastocyst
T925	1484-1489	NN	denotes	stage
T929	1489-1490	.	denotes	.
T930	1490-1643	sentence	denotes	This establishes that μ-calpain and m-calpain have distinct functions, and that m-calpain is vital for development of the preimplantation murine embryo.
T931	1491-1495	DT	denotes	This
T932	1496-1507	VBZ	denotes	establishes
T933	1508-1512	IN	denotes	that
T935	1513-1514	NN	denotes	μ
T937	1514-1515	HYPH	denotes	-
T936	1515-1522	NN	denotes	calpain
T938	1523-1526	CC	denotes	and
T939	1527-1528	NN	denotes	m
T941	1528-1529	HYPH	denotes	-
T940	1529-1536	NN	denotes	calpain
T934	1537-1541	VBP	denotes	have
T942	1542-1550	JJ	denotes	distinct
T943	1551-1560	NNS	denotes	functions
T944	1560-1562	,	denotes	,
T945	1562-1565	CC	denotes	and
T946	1566-1570	IN	denotes	that
T948	1571-1572	NN	denotes	m
T950	1572-1573	HYPH	denotes	-
T949	1573-1580	NN	denotes	calpain
T947	1581-1583	VBZ	denotes	is
T951	1584-1589	JJ	denotes	vital
T952	1590-1593	IN	denotes	for
T953	1594-1605	NN	denotes	development
T954	1606-1608	IN	denotes	of
T955	1609-1612	DT	denotes	the
T957	1613-1628	NN	denotes	preimplantation
T958	1629-1635	JJ	denotes	murine
T956	1636-1642	NN	denotes	embryo
T959	1642-1643	.	denotes	.
T3244	1656-1659	DT	denotes	The
T3246	1660-1663	CD	denotes	two
T3247	1664-1674	JJ	denotes	ubiquitous
T3248	1675-1678	NN	denotes	Ca2
T3250	1678-1679	SYM	denotes	+
T3251	1679-1680	HYPH	denotes	-
T3249	1680-1689	JJ	denotes	dependent
T3252	1689-1691	,	denotes	,
T3253	1691-1699	NN	denotes	cysteine
T3245	1700-1709	NNS	denotes	proteases
T3255	1710-1715	VBN	denotes	known
T3256	1716-1718	IN	denotes	as
T3257	1719-1720	NN	denotes	μ
T3259	1720-1721	HYPH	denotes	-
T3258	1721-1728	NN	denotes	calpain
T3260	1729-1730	-LRB-	denotes	(
T3261	1730-1737	NN	denotes	calpain
T3262	1737-1738	HYPH	denotes	-
T3263	1738-1739	CD	denotes	1
T3264	1739-1740	-RRB-	denotes	)
T3265	1741-1744	CC	denotes	and
T3266	1745-1746	NN	denotes	m
T3268	1746-1747	HYPH	denotes	-
T3267	1747-1754	NN	denotes	calpain
T3269	1755-1756	-LRB-	denotes	(
T3270	1756-1762	NN	denotes	capain
T3271	1762-1763	HYPH	denotes	-
T3272	1763-1764	CD	denotes	2
T3273	1764-1765	-RRB-	denotes	)
T3274	1765-1767	,	denotes	,
T3254	1767-1770	VBP	denotes	are
T3275	1771-1774	DT	denotes	the
T3277	1775-1783	NN	denotes	founding
T3276	1784-1791	NNS	denotes	members
T3278	1792-1794	IN	denotes	of
T3279	1795-1796	DT	denotes	a
T3281	1797-1801	NN	denotes	gene
T3280	1802-1808	NN	denotes	family
T3282	1809-1819	VBG	denotes	comprising
T3283	1820-1822	CD	denotes	13
T3284	1823-1828	NNS	denotes	genes
T3285	1829-1831	IN	denotes	in
T3286	1832-1839	NNS	denotes	mammals
T3287	1840-1841	-LRB-	denotes	[
T3288	1841-1842	CD	denotes	1
T3289	1842-1843	SYM	denotes	-
T3290	1843-1844	CD	denotes	3
T3291	1844-1845	-RRB-	denotes	]
T3292	1845-1846	.	denotes	.
T3293	1846-2079	sentence	denotes	Both are heterodimeric enzymes consisting of distinct 80 kDa catalytic subunits, encoded by the Capn1 (μ-80 k) and Capn2 (m-80 k) genes, respectively, that associate with a common 28 kDa regulatory subunit encoded by the Capn4 gene.
T3294	1847-1851	DT	denotes	Both
T3295	1852-1855	VBP	denotes	are
T3296	1856-1869	JJ	denotes	heterodimeric
T3297	1870-1877	NNS	denotes	enzymes
T3298	1878-1888	VBG	denotes	consisting
T3299	1889-1891	IN	denotes	of
T3300	1892-1900	JJ	denotes	distinct
T3302	1901-1903	CD	denotes	80
T3303	1904-1907	NN	denotes	kDa
T3304	1908-1917	JJ	denotes	catalytic
T3301	1918-1926	NNS	denotes	subunits
T3305	1926-1928	,	denotes	,
T3306	1928-1935	VBN	denotes	encoded
T3307	1936-1938	IN	denotes	by
T3308	1939-1942	DT	denotes	the
T3310	1943-1948	NN	denotes	Capn1
T3311	1949-1950	-LRB-	denotes	(
T3313	1950-1951	NN	denotes	μ
T3314	1951-1952	HYPH	denotes	-
T3315	1952-1954	CD	denotes	80
T3312	1955-1956	NN	denotes	k
T3316	1956-1957	-RRB-	denotes	)
T3317	1958-1961	CC	denotes	and
T3318	1962-1967	NN	denotes	Capn2
T3319	1968-1969	-LRB-	denotes	(
T3321	1969-1970	NN	denotes	m
T3322	1970-1971	HYPH	denotes	-
T3323	1971-1973	CD	denotes	80
T3320	1974-1975	NN	denotes	k
T3324	1975-1976	-RRB-	denotes	)
T3309	1977-1982	NNS	denotes	genes
T3325	1982-1984	,	denotes	,
T3326	1984-1996	RB	denotes	respectively
T3327	1996-1998	,	denotes	,
T3328	1998-2002	WDT	denotes	that
T3329	2003-2012	VBP	denotes	associate
T3330	2013-2017	IN	denotes	with
T3331	2018-2019	DT	denotes	a
T3333	2020-2026	JJ	denotes	common
T3334	2027-2029	CD	denotes	28
T3335	2030-2033	NN	denotes	kDa
T3336	2034-2044	JJ	denotes	regulatory
T3332	2045-2052	NN	denotes	subunit
T3337	2053-2060	VBN	denotes	encoded
T3338	2061-2063	IN	denotes	by
T3339	2064-2067	DT	denotes	the
T3341	2068-2073	NN	denotes	Capn4
T3340	2074-2078	NN	denotes	gene
T3342	2078-2079	.	denotes	.
T3343	2079-2249	sentence	denotes	The μ-80 k and m-80 k subunits share 62% amino acid sequence identity, and are very similar in terms of structure, protein chemistry, and in vitro substrate specificity.
T3344	2080-2083	DT	denotes	The
T3346	2084-2085	NN	denotes	μ
T3348	2085-2086	HYPH	denotes	-
T3349	2086-2088	CD	denotes	80
T3347	2089-2090	NN	denotes	k
T3350	2091-2094	CC	denotes	and
T3351	2095-2096	NN	denotes	m
T3353	2096-2097	HYPH	denotes	-
T3354	2097-2099	CD	denotes	80
T3352	2100-2101	NN	denotes	k
T3345	2102-2110	NNS	denotes	subunits
T3355	2111-2116	VBP	denotes	share
T3356	2117-2119	CD	denotes	62
T3357	2119-2120	NN	denotes	%
T3359	2121-2126	NN	denotes	amino
T3360	2127-2131	NN	denotes	acid
T3361	2132-2140	NN	denotes	sequence
T3358	2141-2149	NN	denotes	identity
T3362	2149-2151	,	denotes	,
T3363	2151-2154	CC	denotes	and
T3364	2155-2158	VBP	denotes	are
T3365	2159-2163	RB	denotes	very
T3366	2164-2171	JJ	denotes	similar
T3367	2172-2174	IN	denotes	in
T3368	2175-2180	NNS	denotes	terms
T3369	2181-2183	IN	denotes	of
T3370	2184-2193	NN	denotes	structure
T3371	2193-2195	,	denotes	,
T3372	2195-2202	NN	denotes	protein
T3373	2203-2212	NN	denotes	chemistry
T3374	2212-2214	,	denotes	,
T3375	2214-2217	CC	denotes	and
T3376	2218-2220	FW	denotes	in
T3377	2221-2226	FW	denotes	vitro
T3379	2227-2236	NN	denotes	substrate
T3378	2237-2248	NN	denotes	specificity
T3380	2248-2249	.	denotes	.
T3381	2249-2419	sentence	denotes	Despite these similarities, the differential expression patterns of μ- and m-calpain in mammalian tissues suggest they have some isoform specific and distinct functions.
T3382	2250-2257	IN	denotes	Despite
T3384	2258-2263	DT	denotes	these
T3385	2264-2276	NNS	denotes	similarities
T3386	2276-2278	,	denotes	,
T3387	2278-2281	DT	denotes	the
T3389	2282-2294	JJ	denotes	differential
T3390	2295-2305	NN	denotes	expression
T3388	2306-2314	NNS	denotes	patterns
T3391	2315-2317	IN	denotes	of
T3392	2318-2319	NN	denotes	μ
T3394	2319-2320	HYPH	denotes	-
T3395	2321-2324	CC	denotes	and
T3396	2325-2326	NN	denotes	m
T3397	2326-2327	HYPH	denotes	-
T3393	2327-2334	NN	denotes	calpain
T3398	2335-2337	IN	denotes	in
T3399	2338-2347	JJ	denotes	mammalian
T3400	2348-2355	NNS	denotes	tissues
T3383	2356-2363	VBP	denotes	suggest
T3401	2364-2368	PRP	denotes	they
T3402	2369-2373	VBP	denotes	have
T3403	2374-2378	DT	denotes	some
T3405	2379-2386	NN	denotes	isoform
T3406	2387-2395	JJ	denotes	specific
T3407	2396-2399	CC	denotes	and
T3408	2400-2408	JJ	denotes	distinct
T3404	2409-2418	NNS	denotes	functions
T3409	2418-2419	.	denotes	.
T3410	2419-2581	sentence	denotes	The μ and m designations derive from the levels of Ca2+ required in vitro for optimal activation; 10–50 μM Ca2+ for μ-calpain and 0.3–0.35 mM Ca2+ for m-calpain.
T3411	2420-2423	DT	denotes	The
T3413	2424-2425	NN	denotes	μ
T3414	2426-2429	CC	denotes	and
T3415	2430-2431	NN	denotes	m
T3412	2432-2444	NNS	denotes	designations
T3416	2445-2451	VBP	denotes	derive
T3417	2452-2456	IN	denotes	from
T3418	2457-2460	DT	denotes	the
T3419	2461-2467	NNS	denotes	levels
T3420	2468-2470	IN	denotes	of
T3421	2471-2474	NN	denotes	Ca2
T3422	2474-2475	SYM	denotes	+
T3423	2476-2484	VBN	denotes	required
T3424	2485-2487	FW	denotes	in
T3425	2488-2493	FW	denotes	vitro
T3426	2494-2497	IN	denotes	for
T3427	2498-2505	JJ	denotes	optimal
T3428	2506-2516	NN	denotes	activation
T3429	2516-2517	:	denotes	;
T3430	2518-2520	CD	denotes	10
T3432	2520-2521	SYM	denotes	–
T3431	2521-2523	CD	denotes	50
T3433	2524-2526	NN	denotes	μM
T3434	2527-2530	NN	denotes	Ca2
T3435	2530-2531	SYM	denotes	+
T3436	2532-2535	IN	denotes	for
T3437	2536-2537	NN	denotes	μ
T3439	2537-2538	HYPH	denotes	-
T3438	2538-2545	NN	denotes	calpain
T3440	2546-2549	CC	denotes	and
T3441	2550-2553	CD	denotes	0.3
T3443	2553-2554	SYM	denotes	–
T3442	2554-2558	CD	denotes	0.35
T3444	2559-2561	NN	denotes	mM
T3445	2562-2565	NN	denotes	Ca2
T3446	2565-2566	SYM	denotes	+
T3447	2567-2570	IN	denotes	for
T3448	2571-2572	NN	denotes	m
T3450	2572-2573	HYPH	denotes	-
T3449	2573-2580	NN	denotes	calpain
T3451	2580-2581	.	denotes	.
T3452	2581-2743	sentence	denotes	It is generally assumed that μ- and m-calpain maintain their differential sensitivities to calcium in vivo, although this has not yet been strictly demonstrated.
T3453	2582-2584	PRP	denotes	It
T3455	2585-2587	VBZ	denotes	is
T3456	2588-2597	RB	denotes	generally
T3454	2598-2605	VBN	denotes	assumed
T3457	2606-2610	IN	denotes	that
T3459	2611-2612	NN	denotes	μ
T3461	2612-2613	HYPH	denotes	-
T3462	2614-2617	CC	denotes	and
T3463	2618-2619	NN	denotes	m
T3464	2619-2620	HYPH	denotes	-
T3460	2620-2627	NN	denotes	calpain
T3458	2628-2636	VBP	denotes	maintain
T3465	2637-2642	PRP$	denotes	their
T3467	2643-2655	JJ	denotes	differential
T3466	2656-2669	NNS	denotes	sensitivities
T3468	2670-2672	IN	denotes	to
T3469	2673-2680	NN	denotes	calcium
T3470	2681-2683	FW	denotes	in
T3471	2684-2688	FW	denotes	vivo
T3472	2688-2690	,	denotes	,
T3473	2690-2698	IN	denotes	although
T3475	2699-2703	DT	denotes	this
T3476	2704-2707	VBZ	denotes	has
T3477	2708-2711	RB	denotes	not
T3478	2712-2715	RB	denotes	yet
T3479	2716-2720	VBN	denotes	been
T3480	2721-2729	RB	denotes	strictly
T3474	2730-2742	VBN	denotes	demonstrated
T3481	2742-2743	.	denotes	.
T3482	2743-2929	sentence	denotes	Furthermore, since the cytoplasmic free Ca2+ concentration is typically less than 1 μM, it is also assumed that other in vivo factors must contribute to regulation of these enzymes [3].
T3483	2744-2755	RB	denotes	Furthermore
T3485	2755-2757	,	denotes	,
T3486	2757-2762	IN	denotes	since
T3488	2763-2766	DT	denotes	the
T3490	2767-2778	JJ	denotes	cytoplasmic
T3491	2779-2783	JJ	denotes	free
T3492	2784-2787	NN	denotes	Ca2
T3493	2787-2788	SYM	denotes	+
T3489	2789-2802	NN	denotes	concentration
T3487	2803-2805	VBZ	denotes	is
T3494	2806-2815	RB	denotes	typically
T3495	2816-2820	JJR	denotes	less
T3497	2821-2825	IN	denotes	than
T3496	2826-2827	CD	denotes	1
T3498	2828-2830	NN	denotes	μM
T3499	2830-2832	,	denotes	,
T3500	2832-2834	PRP	denotes	it
T3501	2835-2837	VBZ	denotes	is
T3502	2838-2842	RB	denotes	also
T3484	2843-2850	VBN	denotes	assumed
T3503	2851-2855	IN	denotes	that
T3505	2856-2861	JJ	denotes	other
T3507	2862-2864	FW	denotes	in
T3508	2865-2869	FW	denotes	vivo
T3506	2870-2877	NNS	denotes	factors
T3509	2878-2882	MD	denotes	must
T3504	2883-2893	VB	denotes	contribute
T3510	2894-2896	IN	denotes	to
T3511	2897-2907	NN	denotes	regulation
T3512	2908-2910	IN	denotes	of
T3513	2911-2916	DT	denotes	these
T3514	2917-2924	NNS	denotes	enzymes
T3515	2925-2926	-LRB-	denotes	[
T3516	2926-2927	CD	denotes	3
T3517	2927-2928	-RRB-	denotes	]
T3518	2928-2929	.	denotes	.
T3519	2929-3099	sentence	denotes	Without apriori knowledge of the factors regulating calpain activity or their relevant substrates, elucidation of biological functions for calpains presents a challenge.
T3520	2930-2937	IN	denotes	Without
T3522	2938-2945	FW	denotes	apriori
T3523	2946-2955	NN	denotes	knowledge
T3524	2956-2958	IN	denotes	of
T3525	2959-2962	DT	denotes	the
T3526	2963-2970	NNS	denotes	factors
T3527	2971-2981	VBG	denotes	regulating
T3528	2982-2989	NN	denotes	calpain
T3529	2990-2998	NN	denotes	activity
T3530	2999-3001	CC	denotes	or
T3531	3002-3007	PRP$	denotes	their
T3533	3008-3016	JJ	denotes	relevant
T3532	3017-3027	NNS	denotes	substrates
T3534	3027-3029	,	denotes	,
T3535	3029-3040	NN	denotes	elucidation
T3536	3041-3043	IN	denotes	of
T3537	3044-3054	JJ	denotes	biological
T3538	3055-3064	NNS	denotes	functions
T3539	3065-3068	IN	denotes	for
T3540	3069-3077	NNS	denotes	calpains
T3521	3078-3086	VBZ	denotes	presents
T3541	3087-3088	DT	denotes	a
T3542	3089-3098	NN	denotes	challenge
T3543	3098-3099	.	denotes	.
T3544	3099-3514	sentence	denotes	Research on calpains has linked them with a wide variety of functions including muscle growth, development, degeneration (3), neuronal growth and neurodegeneration [4], cell cycle progression [5,6], signal cascades triggered by integrins and growth factors [7], membrane protrusion [8], remodeling of the cytoskeleton and cell migration [9-15], and regulation of cell death via both necrosis and apoptosis [16-22].
T3545	3100-3108	NN	denotes	Research
T3547	3109-3111	IN	denotes	on
T3548	3112-3120	NNS	denotes	calpains
T3549	3121-3124	VBZ	denotes	has
T3546	3125-3131	VBN	denotes	linked
T3550	3132-3136	PRP	denotes	them
T3551	3137-3141	IN	denotes	with
T3552	3142-3143	DT	denotes	a
T3554	3144-3148	JJ	denotes	wide
T3553	3149-3156	NN	denotes	variety
T3555	3157-3159	IN	denotes	of
T3556	3160-3169	NNS	denotes	functions
T3557	3170-3179	VBG	denotes	including
T3558	3180-3186	NN	denotes	muscle
T3559	3187-3193	NN	denotes	growth
T3560	3193-3195	,	denotes	,
T3561	3195-3206	NN	denotes	development
T3562	3206-3208	,	denotes	,
T3563	3208-3220	NN	denotes	degeneration
T3564	3221-3222	-LRB-	denotes	(
T3565	3222-3223	CD	denotes	3
T3566	3223-3224	-RRB-	denotes	)
T3567	3224-3226	,	denotes	,
T3568	3226-3234	JJ	denotes	neuronal
T3569	3235-3241	NN	denotes	growth
T3570	3242-3245	CC	denotes	and
T3571	3246-3263	NN	denotes	neurodegeneration
T3572	3264-3265	-LRB-	denotes	[
T3573	3265-3266	CD	denotes	4
T3574	3266-3267	-RRB-	denotes	]
T3575	3267-3269	,	denotes	,
T3576	3269-3273	NN	denotes	cell
T3577	3274-3279	NN	denotes	cycle
T3578	3280-3291	NN	denotes	progression
T3579	3292-3293	-LRB-	denotes	[
T3581	3293-3294	CD	denotes	5
T3582	3294-3295	,	denotes	,
T3580	3295-3296	CD	denotes	6
T3583	3296-3297	-RRB-	denotes	]
T3584	3297-3299	,	denotes	,
T3585	3299-3305	NN	denotes	signal
T3586	3306-3314	NNS	denotes	cascades
T3587	3315-3324	VBN	denotes	triggered
T3588	3325-3327	IN	denotes	by
T3589	3328-3337	NNS	denotes	integrins
T3590	3338-3341	CC	denotes	and
T3591	3342-3348	NN	denotes	growth
T3592	3349-3356	NNS	denotes	factors
T3593	3357-3358	-LRB-	denotes	[
T3594	3358-3359	CD	denotes	7
T3595	3359-3360	-RRB-	denotes	]
T3596	3360-3362	,	denotes	,
T3597	3362-3370	NN	denotes	membrane
T3598	3371-3381	NN	denotes	protrusion
T3599	3382-3383	-LRB-	denotes	[
T3600	3383-3384	CD	denotes	8
T3601	3384-3385	-RRB-	denotes	]
T3602	3385-3387	,	denotes	,
T3603	3387-3397	VBG	denotes	remodeling
T3604	3398-3400	IN	denotes	of
T3605	3401-3404	DT	denotes	the
T3606	3405-3417	NN	denotes	cytoskeleton
T3607	3418-3421	CC	denotes	and
T3608	3422-3426	NN	denotes	cell
T3609	3427-3436	NN	denotes	migration
T3610	3437-3438	-LRB-	denotes	[
T3611	3438-3439	CD	denotes	9
T3612	3439-3440	SYM	denotes	-
T3613	3440-3442	CD	denotes	15
T3614	3442-3443	-RRB-	denotes	]
T3615	3443-3445	,	denotes	,
T3616	3445-3448	CC	denotes	and
T3617	3449-3459	NN	denotes	regulation
T3618	3460-3462	IN	denotes	of
T3619	3463-3467	NN	denotes	cell
T3620	3468-3473	NN	denotes	death
T3621	3474-3477	IN	denotes	via
T3622	3478-3482	CC	denotes	both
T3623	3483-3491	NN	denotes	necrosis
T3624	3492-3495	CC	denotes	and
T3625	3496-3505	NN	denotes	apoptosis
T3626	3506-3507	-LRB-	denotes	[
T3627	3507-3509	CD	denotes	16
T3628	3509-3510	SYM	denotes	-
T3629	3510-3512	CD	denotes	22
T3630	3512-3513	-RRB-	denotes	]
T3631	3513-3514	.	denotes	.
T3632	3514-3666	sentence	denotes	To date, the literature suggests a complex interplay between caspases and calpains [23,24] and impact of calpain on cell death pathway components [25].
T3633	3515-3517	IN	denotes	To
T3635	3518-3522	NN	denotes	date
T3636	3522-3524	,	denotes	,
T3637	3524-3527	DT	denotes	the
T3638	3528-3538	NN	denotes	literature
T3634	3539-3547	VBZ	denotes	suggests
T3639	3548-3549	DT	denotes	a
T3641	3550-3557	JJ	denotes	complex
T3640	3558-3567	NN	denotes	interplay
T3642	3568-3575	IN	denotes	between
T3643	3576-3584	NNS	denotes	caspases
T3644	3585-3588	CC	denotes	and
T3645	3589-3597	NNS	denotes	calpains
T3646	3598-3599	-LRB-	denotes	[
T3648	3599-3601	CD	denotes	23
T3649	3601-3602	,	denotes	,
T3647	3602-3604	CD	denotes	24
T3650	3604-3605	-RRB-	denotes	]
T3651	3606-3609	CC	denotes	and
T3652	3610-3616	NN	denotes	impact
T3653	3617-3619	IN	denotes	of
T3654	3620-3627	NN	denotes	calpain
T3655	3628-3630	IN	denotes	on
T3656	3631-3635	NN	denotes	cell
T3657	3636-3641	NN	denotes	death
T3659	3642-3649	NN	denotes	pathway
T3658	3650-3660	NNS	denotes	components
T3660	3661-3662	-LRB-	denotes	[
T3661	3662-3664	CD	denotes	25
T3662	3664-3665	-RRB-	denotes	]
T3663	3665-3666	.	denotes	.
T3664	3666-3829	sentence	denotes	The lack of highly specific cell-permeable inhibitors of calpains contributes to the challenge of investigating and defining calpain functions in these processes.
T3665	3667-3670	DT	denotes	The
T3666	3671-3675	NN	denotes	lack
T3668	3676-3678	IN	denotes	of
T3669	3679-3685	RB	denotes	highly
T3670	3686-3694	JJ	denotes	specific
T3672	3695-3699	NN	denotes	cell
T3674	3699-3700	HYPH	denotes	-
T3673	3700-3709	JJ	denotes	permeable
T3671	3710-3720	NNS	denotes	inhibitors
T3675	3721-3723	IN	denotes	of
T3676	3724-3732	NNS	denotes	calpains
T3667	3733-3744	VBZ	denotes	contributes
T3677	3745-3747	IN	denotes	to
T3678	3748-3751	DT	denotes	the
T3679	3752-3761	NN	denotes	challenge
T3680	3762-3764	IN	denotes	of
T3681	3765-3778	VBG	denotes	investigating
T3682	3779-3782	CC	denotes	and
T3683	3783-3791	VBG	denotes	defining
T3684	3792-3799	NN	denotes	calpain
T3685	3800-3809	NNS	denotes	functions
T3686	3810-3812	IN	denotes	in
T3687	3813-3818	DT	denotes	these
T3688	3819-3828	NNS	denotes	processes
T3689	3828-3829	.	denotes	.
T3690	3829-4036	sentence	denotes	Although over-expression of calpastatin, the endogenous protein inhibitor of μ- and m-calpain provides an important approach for these efforts, it will not distinguish isoform specific functions [24,26,27].
T3691	3830-3838	IN	denotes	Although
T3693	3839-3854	NN	denotes	over-expression
T3694	3855-3857	IN	denotes	of
T3695	3858-3869	NN	denotes	calpastatin
T3696	3869-3871	,	denotes	,
T3697	3871-3874	DT	denotes	the
T3699	3875-3885	JJ	denotes	endogenous
T3700	3886-3893	NN	denotes	protein
T3698	3894-3903	NN	denotes	inhibitor
T3701	3904-3906	IN	denotes	of
T3702	3907-3908	NN	denotes	μ
T3704	3908-3909	HYPH	denotes	-
T3705	3910-3913	CC	denotes	and
T3706	3914-3915	NN	denotes	m
T3707	3915-3916	HYPH	denotes	-
T3703	3916-3923	NN	denotes	calpain
T3692	3924-3932	VBZ	denotes	provides
T3709	3933-3935	DT	denotes	an
T3711	3936-3945	JJ	denotes	important
T3710	3946-3954	NN	denotes	approach
T3712	3955-3958	IN	denotes	for
T3713	3959-3964	DT	denotes	these
T3714	3965-3972	NNS	denotes	efforts
T3715	3972-3974	,	denotes	,
T3716	3974-3976	PRP	denotes	it
T3717	3977-3981	MD	denotes	will
T3718	3982-3985	RB	denotes	not
T3708	3986-3997	VB	denotes	distinguish
T3719	3998-4005	NN	denotes	isoform
T3720	4006-4014	JJ	denotes	specific
T3721	4015-4024	NNS	denotes	functions
T3722	4025-4026	-LRB-	denotes	[
T3724	4026-4028	CD	denotes	24
T3725	4028-4029	,	denotes	,
T3726	4029-4031	CD	denotes	26
T3727	4031-4032	,	denotes	,
T3723	4032-4034	CD	denotes	27
T3728	4034-4035	-RRB-	denotes	]
T3729	4035-4036	.	denotes	.
T3730	4036-4276	sentence	denotes	Some work has suggested isoform specific roles, such as a role for m-calpain in epidermal growth factor (EGF)-induced cell motility [28,29] and a role for μ-calpain in interferon-inducible protein 9-induced migration of keratinocytes [28].
T3731	4037-4041	DT	denotes	Some
T3732	4042-4046	NN	denotes	work
T3734	4047-4050	VBZ	denotes	has
T3733	4051-4060	VBN	denotes	suggested
T3735	4061-4068	NN	denotes	isoform
T3736	4069-4077	JJ	denotes	specific
T3737	4078-4083	NNS	denotes	roles
T3738	4083-4085	,	denotes	,
T3739	4085-4089	JJ	denotes	such
T3740	4090-4092	IN	denotes	as
T3741	4093-4094	DT	denotes	a
T3742	4095-4099	NN	denotes	role
T3743	4100-4103	IN	denotes	for
T3744	4104-4105	NN	denotes	m
T3746	4105-4106	HYPH	denotes	-
T3745	4106-4113	NN	denotes	calpain
T3747	4114-4116	IN	denotes	in
T3748	4117-4126	JJ	denotes	epidermal
T3750	4127-4133	NN	denotes	growth
T3749	4134-4140	NN	denotes	factor
T3752	4141-4142	-LRB-	denotes	(
T3753	4142-4145	NN	denotes	EGF
T3754	4145-4146	-RRB-	denotes	)
T3755	4146-4147	HYPH	denotes	-
T3751	4147-4154	VBN	denotes	induced
T3757	4155-4159	NN	denotes	cell
T3756	4160-4168	NN	denotes	motility
T3758	4169-4170	-LRB-	denotes	[
T3760	4170-4172	CD	denotes	28
T3761	4172-4173	,	denotes	,
T3759	4173-4175	CD	denotes	29
T3762	4175-4176	-RRB-	denotes	]
T3763	4177-4180	CC	denotes	and
T3764	4181-4182	DT	denotes	a
T3765	4183-4187	NN	denotes	role
T3766	4188-4191	IN	denotes	for
T3767	4192-4193	NN	denotes	μ
T3769	4193-4194	HYPH	denotes	-
T3768	4194-4201	NN	denotes	calpain
T3770	4202-4204	IN	denotes	in
T3771	4205-4215	NN	denotes	interferon
T3773	4215-4216	HYPH	denotes	-
T3772	4216-4225	JJ	denotes	inducible
T3775	4226-4233	NN	denotes	protein
T3777	4234-4235	CD	denotes	9
T3778	4235-4236	HYPH	denotes	-
T3776	4236-4243	VBN	denotes	induced
T3774	4244-4253	NN	denotes	migration
T3779	4254-4256	IN	denotes	of
T3780	4257-4270	NNS	denotes	keratinocytes
T3781	4271-4272	-LRB-	denotes	[
T3782	4272-4274	CD	denotes	28
T3783	4274-4275	-RRB-	denotes	]
T3784	4275-4276	.	denotes	.
T3785	4276-4461	sentence	denotes	A cell permeable calpain inhibitor (which likely inhibits other thiol-proteases as well) has been used to select cells lacking μ-calpain which display reduced proliferation rates [30].
T3786	4277-4278	DT	denotes	A
T3788	4279-4283	NN	denotes	cell
T3789	4284-4293	JJ	denotes	permeable
T3790	4294-4301	NN	denotes	calpain
T3787	4302-4311	NN	denotes	inhibitor
T3792	4312-4313	-LRB-	denotes	(
T3793	4313-4318	WDT	denotes	which
T3795	4319-4325	RB	denotes	likely
T3794	4326-4334	VBZ	denotes	inhibits
T3796	4335-4340	JJ	denotes	other
T3798	4341-4346	NN	denotes	thiol
T3799	4346-4347	HYPH	denotes	-
T3797	4347-4356	NNS	denotes	proteases
T3800	4357-4359	RB	denotes	as
T3801	4360-4364	RB	denotes	well
T3802	4364-4365	-RRB-	denotes	)
T3803	4366-4369	VBZ	denotes	has
T3804	4370-4374	VBN	denotes	been
T3791	4375-4379	VBN	denotes	used
T3805	4380-4382	TO	denotes	to
T3806	4383-4389	VB	denotes	select
T3807	4390-4395	NNS	denotes	cells
T3808	4396-4403	VBG	denotes	lacking
T3809	4404-4405	NN	denotes	μ
T3811	4405-4406	HYPH	denotes	-
T3810	4406-4413	NN	denotes	calpain
T3812	4414-4419	WDT	denotes	which
T3813	4420-4427	VBP	denotes	display
T3814	4428-4435	VBN	denotes	reduced
T3816	4436-4449	NN	denotes	proliferation
T3815	4450-4455	NNS	denotes	rates
T3817	4456-4457	-LRB-	denotes	[
T3818	4457-4459	CD	denotes	30
T3819	4459-4460	-RRB-	denotes	]
T3820	4460-4461	.	denotes	.
T3821	4461-4594	sentence	denotes	Interestingly, m-calpain expression persisted in these cells, suggesting a possible requirement of m-calpain for cell survival [30].
T3822	4462-4475	RB	denotes	Interestingly
T3824	4475-4477	,	denotes	,
T3825	4477-4478	NN	denotes	m
T3827	4478-4479	HYPH	denotes	-
T3826	4479-4486	NN	denotes	calpain
T3828	4487-4497	NN	denotes	expression
T3823	4498-4507	VBD	denotes	persisted
T3829	4508-4510	IN	denotes	in
T3830	4511-4516	DT	denotes	these
T3831	4517-4522	NNS	denotes	cells
T3832	4522-4524	,	denotes	,
T3833	4524-4534	VBG	denotes	suggesting
T3834	4535-4536	DT	denotes	a
T3836	4537-4545	JJ	denotes	possible
T3835	4546-4557	NN	denotes	requirement
T3837	4558-4560	IN	denotes	of
T3838	4561-4562	NN	denotes	m
T3840	4562-4563	HYPH	denotes	-
T3839	4563-4570	NN	denotes	calpain
T3841	4571-4574	IN	denotes	for
T3842	4575-4579	NN	denotes	cell
T3843	4580-4588	NN	denotes	survival
T3844	4589-4590	-LRB-	denotes	[
T3845	4590-4592	CD	denotes	30
T3846	4592-4593	-RRB-	denotes	]
T3847	4593-4594	.	denotes	.
T3848	4594-4779	sentence	denotes	Targeted gene deletion in mice provides a powerful approach to determining the physiological roles of μ- and m-calpain and the opportunity to approach their isoform specific functions.
T3849	4595-4603	VBN	denotes	Targeted
T3851	4604-4608	NN	denotes	gene
T3850	4609-4617	NN	denotes	deletion
T3853	4618-4620	IN	denotes	in
T3854	4621-4625	NNS	denotes	mice
T3852	4626-4634	VBZ	denotes	provides
T3855	4635-4636	DT	denotes	a
T3857	4637-4645	JJ	denotes	powerful
T3856	4646-4654	NN	denotes	approach
T3858	4655-4657	IN	denotes	to
T3859	4658-4669	VBG	denotes	determining
T3860	4670-4673	DT	denotes	the
T3862	4674-4687	JJ	denotes	physiological
T3861	4688-4693	NNS	denotes	roles
T3863	4694-4696	IN	denotes	of
T3864	4697-4698	NN	denotes	μ
T3866	4698-4699	HYPH	denotes	-
T3867	4700-4703	CC	denotes	and
T3868	4704-4705	NN	denotes	m
T3869	4705-4706	HYPH	denotes	-
T3865	4706-4713	NN	denotes	calpain
T3870	4714-4717	CC	denotes	and
T3871	4718-4721	DT	denotes	the
T3872	4722-4733	NN	denotes	opportunity
T3873	4734-4736	TO	denotes	to
T3874	4737-4745	VB	denotes	approach
T3875	4746-4751	PRP$	denotes	their
T3877	4752-4759	NN	denotes	isoform
T3878	4760-4768	JJ	denotes	specific
T3876	4769-4778	NNS	denotes	functions
T3879	4778-4779	.	denotes	.
T3880	4779-4917	sentence	denotes	Initial studies targeted Capn4 based on the prediction that loss of this calpain subunit would abolish activity of both μ- and m-calpain.
T3881	4780-4787	JJ	denotes	Initial
T3882	4788-4795	NNS	denotes	studies
T3883	4796-4804	VBD	denotes	targeted
T3884	4805-4810	NN	denotes	Capn4
T3885	4811-4816	VBN	denotes	based
T3886	4817-4819	IN	denotes	on
T3887	4820-4823	DT	denotes	the
T3888	4824-4834	NN	denotes	prediction
T3889	4835-4839	IN	denotes	that
T3891	4840-4844	NN	denotes	loss
T3892	4845-4847	IN	denotes	of
T3893	4848-4852	DT	denotes	this
T3895	4853-4860	NN	denotes	calpain
T3894	4861-4868	NN	denotes	subunit
T3896	4869-4874	MD	denotes	would
T3890	4875-4882	VB	denotes	abolish
T3897	4883-4891	NN	denotes	activity
T3898	4892-4894	IN	denotes	of
T3899	4895-4899	CC	denotes	both
T3900	4900-4901	NN	denotes	μ
T3902	4901-4902	HYPH	denotes	-
T3903	4903-4906	CC	denotes	and
T3904	4907-4908	NN	denotes	m
T3905	4908-4909	HYPH	denotes	-
T3901	4909-4916	NN	denotes	calpain
T3906	4916-4917	.	denotes	.
T3907	4917-5070	sentence	denotes	Capn4-/- murine embryos died between days 10 and 11 of gestation, and there was no detectable μ- or m-calpain activity in these or younger embryos [31].
T3908	4918-4923	NN	denotes	Capn4
T3910	4923-4924	SYM	denotes	-
T3911	4924-4925	HYPH	denotes	/
T3912	4925-4926	SYM	denotes	-
T3913	4927-4933	JJ	denotes	murine
T3909	4934-4941	NNS	denotes	embryos
T3914	4942-4946	VBD	denotes	died
T3915	4947-4954	IN	denotes	between
T3916	4955-4959	NNS	denotes	days
T3917	4960-4962	CD	denotes	10
T3918	4963-4966	CC	denotes	and
T3919	4967-4969	CD	denotes	11
T3920	4970-4972	IN	denotes	of
T3921	4973-4982	NN	denotes	gestation
T3922	4982-4984	,	denotes	,
T3923	4984-4987	CC	denotes	and
T3924	4988-4993	EX	denotes	there
T3925	4994-4997	VBD	denotes	was
T3926	4998-5000	DT	denotes	no
T3928	5001-5011	JJ	denotes	detectable
T3929	5012-5013	NN	denotes	μ
T3931	5013-5014	HYPH	denotes	-
T3932	5015-5017	CC	denotes	or
T3933	5018-5019	NN	denotes	m
T3934	5019-5020	HYPH	denotes	-
T3930	5020-5027	NN	denotes	calpain
T3927	5028-5036	NN	denotes	activity
T3935	5037-5039	IN	denotes	in
T3936	5040-5045	DT	denotes	these
T3937	5046-5048	CC	denotes	or
T3938	5049-5056	JJR	denotes	younger
T3939	5057-5064	NNS	denotes	embryos
T3940	5065-5066	-LRB-	denotes	[
T3941	5066-5068	CD	denotes	31
T3942	5068-5069	-RRB-	denotes	]
T3943	5069-5070	.	denotes	.
T3944	5070-5403	sentence	denotes	Capn4-/- murine embryonic fibroblasts (MEFs) could be cultured from these embryos, although they also lacked calpain activity as assessed by casein zymography or by the formation of characteristic spectrin breakdown products, and they displayed migration defects consistent with a role for calpain in release of focal adhesions [9].
T3945	5071-5076	NN	denotes	Capn4
T3947	5076-5077	SYM	denotes	-
T3948	5077-5078	HYPH	denotes	/
T3949	5078-5079	SYM	denotes	-
T3950	5080-5086	JJ	denotes	murine
T3952	5087-5096	JJ	denotes	embryonic
T3951	5097-5108	NNS	denotes	fibroblasts
T3953	5109-5110	-LRB-	denotes	(
T3954	5110-5114	NNS	denotes	MEFs
T3955	5114-5115	-RRB-	denotes	)
T3956	5116-5121	MD	denotes	could
T3957	5122-5124	VB	denotes	be
T3946	5125-5133	VBN	denotes	cultured
T3958	5134-5138	IN	denotes	from
T3959	5139-5144	DT	denotes	these
T3960	5145-5152	NNS	denotes	embryos
T3961	5152-5154	,	denotes	,
T3962	5154-5162	IN	denotes	although
T3964	5163-5167	PRP	denotes	they
T3965	5168-5172	RB	denotes	also
T3963	5173-5179	VBD	denotes	lacked
T3966	5180-5187	NN	denotes	calpain
T3967	5188-5196	NN	denotes	activity
T3968	5197-5199	IN	denotes	as
T3969	5200-5208	VBN	denotes	assessed
T3970	5209-5211	IN	denotes	by
T3971	5212-5218	NN	denotes	casein
T3972	5219-5229	NN	denotes	zymography
T3973	5230-5232	CC	denotes	or
T3974	5233-5235	IN	denotes	by
T3975	5236-5239	DT	denotes	the
T3976	5240-5249	NN	denotes	formation
T3977	5250-5252	IN	denotes	of
T3978	5253-5267	JJ	denotes	characteristic
T3980	5268-5276	NN	denotes	spectrin
T3981	5277-5286	NN	denotes	breakdown
T3979	5287-5295	NNS	denotes	products
T3982	5295-5297	,	denotes	,
T3983	5297-5300	CC	denotes	and
T3984	5301-5305	PRP	denotes	they
T3985	5306-5315	VBD	denotes	displayed
T3986	5316-5325	NN	denotes	migration
T3987	5326-5333	NNS	denotes	defects
T3988	5334-5344	JJ	denotes	consistent
T3989	5345-5349	IN	denotes	with
T3990	5350-5351	DT	denotes	a
T3991	5352-5356	NN	denotes	role
T3992	5357-5360	IN	denotes	for
T3993	5361-5368	NN	denotes	calpain
T3994	5369-5371	IN	denotes	in
T3995	5372-5379	NN	denotes	release
T3996	5380-5382	IN	denotes	of
T3997	5383-5388	JJ	denotes	focal
T3998	5389-5398	NNS	denotes	adhesions
T3999	5399-5400	-LRB-	denotes	[
T4000	5400-5401	CD	denotes	9
T4001	5401-5402	-RRB-	denotes	]
T4002	5402-5403	.	denotes	.
T4003	5403-5582	sentence	denotes	An independently derived Capn4 knockout, involving a more extensive deletion of the gene, resulted in an earlier embryonic lethality, apparently at a pre-implantation stage [32].
T4004	5404-5406	DT	denotes	An
T4006	5407-5420	RB	denotes	independently
T4007	5421-5428	VBN	denotes	derived
T4008	5429-5434	NN	denotes	Capn4
T4005	5435-5443	NN	denotes	knockout
T4010	5443-5445	,	denotes	,
T4011	5445-5454	VBG	denotes	involving
T4012	5455-5456	DT	denotes	a
T4014	5457-5461	RBR	denotes	more
T4015	5462-5471	JJ	denotes	extensive
T4013	5472-5480	NN	denotes	deletion
T4016	5481-5483	IN	denotes	of
T4017	5484-5487	DT	denotes	the
T4018	5488-5492	NN	denotes	gene
T4019	5492-5494	,	denotes	,
T4009	5494-5502	VBD	denotes	resulted
T4020	5503-5505	IN	denotes	in
T4021	5506-5508	DT	denotes	an
T4023	5509-5516	JJR	denotes	earlier
T4024	5517-5526	JJ	denotes	embryonic
T4022	5527-5536	NN	denotes	lethality
T4025	5536-5538	,	denotes	,
T4026	5538-5548	RB	denotes	apparently
T4027	5549-5551	IN	denotes	at
T4028	5552-5553	DT	denotes	a
T4030	5554-5570	JJ	denotes	pre-implantation
T4029	5571-5576	NN	denotes	stage
T4031	5577-5578	-LRB-	denotes	[
T4032	5578-5580	CD	denotes	32
T4033	5580-5581	-RRB-	denotes	]
T4034	5581-5582	.	denotes	.
T4035	5582-5899	sentence	denotes	The different times of embryonic lethality suggested that the first reported Capn4-/- mice [31] were targeted with a hypomorphic mutation, which retained some small level of calpain activity, allowing for their survival to mid-gestation, while the second reported Capn4-/- mice [32] represented a true null mutation.
T4036	5583-5586	DT	denotes	The
T4038	5587-5596	JJ	denotes	different
T4037	5597-5602	NNS	denotes	times
T4040	5603-5605	IN	denotes	of
T4041	5606-5615	JJ	denotes	embryonic
T4042	5616-5625	NN	denotes	lethality
T4039	5626-5635	VBD	denotes	suggested
T4043	5636-5640	IN	denotes	that
T4045	5641-5644	DT	denotes	the
T4047	5645-5650	JJ	denotes	first
T4048	5651-5659	VBN	denotes	reported
T4049	5660-5665	NN	denotes	Capn4
T4050	5665-5666	SYM	denotes	-
T4051	5666-5667	HYPH	denotes	/
T4052	5667-5668	SYM	denotes	-
T4046	5669-5673	NNS	denotes	mice
T4053	5674-5675	-LRB-	denotes	[
T4054	5675-5677	CD	denotes	31
T4055	5677-5678	-RRB-	denotes	]
T4056	5679-5683	VBD	denotes	were
T4044	5684-5692	VBN	denotes	targeted
T4057	5693-5697	IN	denotes	with
T4058	5698-5699	DT	denotes	a
T4060	5700-5711	JJ	denotes	hypomorphic
T4059	5712-5720	NN	denotes	mutation
T4061	5720-5722	,	denotes	,
T4062	5722-5727	WDT	denotes	which
T4063	5728-5736	VBD	denotes	retained
T4064	5737-5741	DT	denotes	some
T4066	5742-5747	JJ	denotes	small
T4065	5748-5753	NN	denotes	level
T4067	5754-5756	IN	denotes	of
T4068	5757-5764	NN	denotes	calpain
T4069	5765-5773	NN	denotes	activity
T4070	5773-5775	,	denotes	,
T4071	5775-5783	VBG	denotes	allowing
T4072	5784-5787	IN	denotes	for
T4073	5788-5793	PRP$	denotes	their
T4074	5794-5802	NN	denotes	survival
T4075	5803-5805	IN	denotes	to
T4076	5806-5819	NN	denotes	mid-gestation
T4077	5819-5821	,	denotes	,
T4078	5821-5826	IN	denotes	while
T4080	5827-5830	DT	denotes	the
T4082	5831-5837	JJ	denotes	second
T4083	5838-5846	VBN	denotes	reported
T4084	5847-5852	NN	denotes	Capn4
T4085	5852-5853	SYM	denotes	-
T4086	5853-5854	HYPH	denotes	/
T4087	5854-5855	SYM	denotes	-
T4081	5856-5860	NNS	denotes	mice
T4088	5861-5862	-LRB-	denotes	[
T4089	5862-5864	CD	denotes	32
T4090	5864-5865	-RRB-	denotes	]
T4079	5866-5877	VBD	denotes	represented
T4091	5878-5879	DT	denotes	a
T4093	5880-5884	JJ	denotes	true
T4094	5885-5889	JJ	denotes	null
T4092	5890-5898	NN	denotes	mutation
T4095	5898-5899	.	denotes	.
T4096	5899-6133	sentence	denotes	Disruption of Capn1, encoding the μ-calpain catalytic subunit, was subsequently reported to result in fertile, viable mice with some mild defects in the μ-calpain rich platelets relating to their aggregation and clot retraction [33].
T4097	5900-5910	NN	denotes	Disruption
T4099	5911-5913	IN	denotes	of
T4100	5914-5919	NN	denotes	Capn1
T4101	5919-5921	,	denotes	,
T4102	5921-5929	VBG	denotes	encoding
T4103	5930-5933	DT	denotes	the
T4105	5934-5935	NN	denotes	μ
T4107	5935-5936	HYPH	denotes	-
T4106	5936-5943	NN	denotes	calpain
T4108	5944-5953	JJ	denotes	catalytic
T4104	5954-5961	NN	denotes	subunit
T4109	5961-5963	,	denotes	,
T4110	5963-5966	VBD	denotes	was
T4111	5967-5979	RB	denotes	subsequently
T4098	5980-5988	VBN	denotes	reported
T4112	5989-5991	TO	denotes	to
T4113	5992-5998	VB	denotes	result
T4114	5999-6001	IN	denotes	in
T4115	6002-6009	JJ	denotes	fertile
T4117	6009-6011	,	denotes	,
T4118	6011-6017	JJ	denotes	viable
T4116	6018-6022	NNS	denotes	mice
T4119	6023-6027	IN	denotes	with
T4120	6028-6032	DT	denotes	some
T4122	6033-6037	JJ	denotes	mild
T4121	6038-6045	NNS	denotes	defects
T4123	6046-6048	IN	denotes	in
T4124	6049-6052	DT	denotes	the
T4126	6053-6054	NN	denotes	μ
T4128	6054-6055	HYPH	denotes	-
T4127	6055-6062	NN	denotes	calpain
T4129	6063-6067	JJ	denotes	rich
T4125	6068-6077	NNS	denotes	platelets
T4130	6078-6086	VBG	denotes	relating
T4131	6087-6089	IN	denotes	to
T4132	6090-6095	PRP$	denotes	their
T4133	6096-6107	NN	denotes	aggregation
T4134	6108-6111	CC	denotes	and
T4135	6112-6116	NN	denotes	clot
T4136	6117-6127	NN	denotes	retraction
T4137	6128-6129	-LRB-	denotes	[
T4138	6129-6131	CD	denotes	33
T4139	6131-6132	-RRB-	denotes	]
T4140	6132-6133	.	denotes	.
T4141	6133-6281	sentence	denotes	The fact that Capn4 null mice die during embryogenesis indicates that at least one of the ubiquitous calpains is essential for development to term.
T4142	6134-6137	DT	denotes	The
T4143	6138-6142	NN	denotes	fact
T4145	6143-6147	IN	denotes	that
T4147	6148-6153	NN	denotes	Capn4
T4148	6154-6158	JJ	denotes	null
T4149	6159-6163	NNS	denotes	mice
T4146	6164-6167	VBP	denotes	die
T4150	6168-6174	IN	denotes	during
T4151	6175-6188	NN	denotes	embryogenesis
T4144	6189-6198	VBZ	denotes	indicates
T4152	6199-6203	IN	denotes	that
T4154	6204-6206	RB	denotes	at
T4156	6207-6212	RBS	denotes	least
T4155	6213-6216	CD	denotes	one
T4157	6217-6219	IN	denotes	of
T4158	6220-6223	DT	denotes	the
T4160	6224-6234	JJ	denotes	ubiquitous
T4159	6235-6243	NNS	denotes	calpains
T4153	6244-6246	VBZ	denotes	is
T4161	6247-6256	JJ	denotes	essential
T4162	6257-6260	IN	denotes	for
T4163	6261-6272	NN	denotes	development
T4164	6273-6275	IN	denotes	to
T4165	6276-6280	NN	denotes	term
T4166	6280-6281	.	denotes	.
T4167	6281-6544	sentence	denotes	The viability of Capn1-deficient mice does not however distinguish between two possibilities: either that m-calpain is specifically required during embryogenesis, or that either form of calpain alone is sufficient and can compensate for the absence of the other.
T4168	6282-6285	DT	denotes	The
T4169	6286-6295	NN	denotes	viability
T4171	6296-6298	IN	denotes	of
T4172	6299-6304	NN	denotes	Capn1
T4174	6304-6305	HYPH	denotes	-
T4173	6305-6314	JJ	denotes	deficient
T4175	6315-6319	NNS	denotes	mice
T4176	6320-6324	VBZ	denotes	does
T4177	6325-6328	RB	denotes	not
T4178	6329-6336	RB	denotes	however
T4170	6337-6348	VB	denotes	distinguish
T4179	6349-6356	IN	denotes	between
T4180	6357-6360	CD	denotes	two
T4181	6361-6374	NNS	denotes	possibilities
T4182	6374-6376	:	denotes	:
T4183	6376-6382	CC	denotes	either
T4185	6383-6387	IN	denotes	that
T4186	6388-6389	NN	denotes	m
T4188	6389-6390	HYPH	denotes	-
T4187	6390-6397	NN	denotes	calpain
T4189	6398-6400	VBZ	denotes	is
T4190	6401-6413	RB	denotes	specifically
T4184	6414-6422	VBN	denotes	required
T4191	6423-6429	IN	denotes	during
T4192	6430-6443	NN	denotes	embryogenesis
T4193	6443-6445	,	denotes	,
T4194	6445-6447	CC	denotes	or
T4195	6448-6452	IN	denotes	that
T4197	6453-6459	CC	denotes	either
T4198	6460-6464	NN	denotes	form
T4199	6465-6467	IN	denotes	of
T4200	6468-6475	NN	denotes	calpain
T4201	6476-6481	RB	denotes	alone
T4196	6482-6484	VBZ	denotes	is
T4202	6485-6495	JJ	denotes	sufficient
T4203	6496-6499	CC	denotes	and
T4204	6500-6503	MD	denotes	can
T4205	6504-6514	VB	denotes	compensate
T4206	6515-6518	IN	denotes	for
T4207	6519-6522	DT	denotes	the
T4208	6523-6530	NN	denotes	absence
T4209	6531-6533	IN	denotes	of
T4210	6534-6537	DT	denotes	the
T4211	6538-6543	JJ	denotes	other
T4212	6543-6544	.	denotes	.
T4213	6544-6646	sentence	denotes	To resolve this question, we have now knocked out the Capn2 gene encoding the m-80 k subunit in mice.
T4214	6545-6547	TO	denotes	To
T4215	6548-6555	VB	denotes	resolve
T4217	6556-6560	DT	denotes	this
T4218	6561-6569	NN	denotes	question
T4219	6569-6571	,	denotes	,
T4220	6571-6573	PRP	denotes	we
T4221	6574-6578	VBP	denotes	have
T4222	6579-6582	RB	denotes	now
T4216	6583-6590	VBN	denotes	knocked
T4223	6591-6594	RP	denotes	out
T4224	6595-6598	DT	denotes	the
T4226	6599-6604	NN	denotes	Capn2
T4225	6605-6609	NN	denotes	gene
T4227	6610-6618	VBG	denotes	encoding
T4228	6619-6622	DT	denotes	the
T4230	6623-6624	NN	denotes	m
T4232	6624-6625	HYPH	denotes	-
T4233	6625-6627	CD	denotes	80
T4231	6628-6629	NN	denotes	k
T4229	6630-6637	NN	denotes	subunit
T4234	6638-6640	IN	denotes	in
T4235	6641-6645	NNS	denotes	mice
T4236	6645-6646	.	denotes	.
T4237	6646-6791	sentence	denotes	We report here that Capn2 null embryos died prior to the implantation stage, indicating that m-calpain is indispensable for early embryogenesis.
T4238	6647-6649	PRP	denotes	We
T4239	6650-6656	VBP	denotes	report
T4240	6657-6661	RB	denotes	here
T4241	6662-6666	IN	denotes	that
T4243	6667-6672	NN	denotes	Capn2
T4244	6673-6677	JJ	denotes	null
T4245	6678-6685	NNS	denotes	embryos
T4242	6686-6690	VBD	denotes	died
T4246	6691-6696	JJ	denotes	prior
T4247	6697-6699	IN	denotes	to
T4248	6700-6703	DT	denotes	the
T4250	6704-6716	NN	denotes	implantation
T4249	6717-6722	NN	denotes	stage
T4251	6722-6724	,	denotes	,
T4252	6724-6734	VBG	denotes	indicating
T4253	6735-6739	IN	denotes	that
T4255	6740-6741	NN	denotes	m
T4257	6741-6742	HYPH	denotes	-
T4256	6742-6749	NN	denotes	calpain
T4254	6750-6752	VBZ	denotes	is
T4258	6753-6766	JJ	denotes	indispensable
T4259	6767-6770	IN	denotes	for
T4260	6771-6776	JJ	denotes	early
T4261	6777-6790	NN	denotes	embryogenesis
T4262	6790-6791	.	denotes	.
T4263	6791-6945	sentence	denotes	This role cannot be fulfilled by μ-calpain, which is expressed in embryonic stem (ES) cells [31] and is assumed to be present at this stage of gestation.
T4264	6792-6796	DT	denotes	This
T4265	6797-6801	NN	denotes	role
T4267	6802-6805	MD	denotes	can
T4268	6805-6808	RB	denotes	not
T4269	6809-6811	VB	denotes	be
T4266	6812-6821	VBN	denotes	fulfilled
T4270	6822-6824	IN	denotes	by
T4271	6825-6826	NN	denotes	μ
T4273	6826-6827	HYPH	denotes	-
T4272	6827-6834	NN	denotes	calpain
T4274	6834-6836	,	denotes	,
T4275	6836-6841	WDT	denotes	which
T4277	6842-6844	VBZ	denotes	is
T4276	6845-6854	VBN	denotes	expressed
T4278	6855-6857	IN	denotes	in
T4279	6858-6867	JJ	denotes	embryonic
T4280	6868-6872	NN	denotes	stem
T4282	6873-6874	-LRB-	denotes	(
T4283	6874-6876	NN	denotes	ES
T4284	6876-6877	-RRB-	denotes	)
T4281	6878-6883	NNS	denotes	cells
T4285	6884-6885	-LRB-	denotes	[
T4286	6885-6887	CD	denotes	31
T4287	6887-6888	-RRB-	denotes	]
T4288	6889-6892	CC	denotes	and
T4289	6893-6895	VBZ	denotes	is
T4290	6896-6903	VBN	denotes	assumed
T4291	6904-6906	TO	denotes	to
T4292	6907-6909	VB	denotes	be
T4293	6910-6917	JJ	denotes	present
T4294	6918-6920	IN	denotes	at
T4295	6921-6925	DT	denotes	this
T4296	6926-6931	NN	denotes	stage
T4297	6932-6934	IN	denotes	of
T4298	6935-6944	NN	denotes	gestation
T4299	6944-6945	.	denotes	.
T4300	6945-7068	sentence	denotes	This demonstrates unequivocally that m-calpain and μ-calpain have distinct physiological roles during early embryogenesis.
T4301	6946-6950	DT	denotes	This
T4302	6951-6963	VBZ	denotes	demonstrates
T4303	6964-6977	RB	denotes	unequivocally
T4304	6978-6982	IN	denotes	that
T4306	6983-6984	NN	denotes	m
T4308	6984-6985	HYPH	denotes	-
T4307	6985-6992	NN	denotes	calpain
T4309	6993-6996	CC	denotes	and
T4310	6997-6998	NN	denotes	μ
T4312	6998-6999	HYPH	denotes	-
T4311	6999-7006	NN	denotes	calpain
T4305	7007-7011	VBP	denotes	have
T4313	7012-7020	JJ	denotes	distinct
T4315	7021-7034	JJ	denotes	physiological
T4314	7035-7040	NNS	denotes	roles
T4316	7041-7047	IN	denotes	during
T4317	7048-7053	JJ	denotes	early
T4318	7054-7067	NN	denotes	embryogenesis
T4319	7067-7068	.	denotes	.
T4832	7079-7088	NN	denotes	Isolation
T4833	7089-7092	CC	denotes	and
T4834	7093-7109	NN	denotes	characterization
T4835	7110-7112	IN	denotes	of
T4836	7113-7118	NN	denotes	Capn2
T4837	7119-7127	VBN	denotes	targeted
T4839	7128-7130	NN	denotes	ES
T4840	7131-7135	NN	denotes	cell
T4838	7136-7142	NNS	denotes	clones
T4841	7142-7266	sentence	denotes	Two independent Capn2+/- ES cell lines, designated ES27 and ES36, were isolated from a screen of 305 drug-resistant clones.
T4842	7143-7146	CD	denotes	Two
T4844	7147-7158	JJ	denotes	independent
T4845	7159-7164	NN	denotes	Capn2
T4846	7164-7165	SYM	denotes	+
T4847	7165-7166	HYPH	denotes	/
T4848	7166-7167	SYM	denotes	-
T4849	7168-7170	NN	denotes	ES
T4850	7171-7175	NN	denotes	cell
T4843	7176-7181	NNS	denotes	lines
T4852	7181-7183	,	denotes	,
T4853	7183-7193	VBN	denotes	designated
T4854	7194-7198	NN	denotes	ES27
T4855	7199-7202	CC	denotes	and
T4856	7203-7207	NN	denotes	ES36
T4857	7207-7209	,	denotes	,
T4858	7209-7213	VBD	denotes	were
T4851	7214-7222	VBN	denotes	isolated
T4859	7223-7227	IN	denotes	from
T4860	7228-7229	DT	denotes	a
T4861	7230-7236	NN	denotes	screen
T4862	7237-7239	IN	denotes	of
T4863	7240-7243	CD	denotes	305
T4865	7244-7248	NN	denotes	drug
T4867	7248-7249	HYPH	denotes	-
T4866	7249-7258	JJ	denotes	resistant
T4864	7259-7265	NNS	denotes	clones
T4868	7265-7266	.	denotes	.
T4869	7266-7373	sentence	denotes	Correct targeting of the Capn2 locus was established both by Southern blot hybridization and PCR analysis.
T4870	7267-7274	JJ	denotes	Correct
T4871	7275-7284	NN	denotes	targeting
T4873	7285-7287	IN	denotes	of
T4874	7288-7291	DT	denotes	the
T4876	7292-7297	NN	denotes	Capn2
T4875	7298-7303	NN	denotes	locus
T4877	7304-7307	VBD	denotes	was
T4872	7308-7319	VBN	denotes	established
T4878	7320-7324	CC	denotes	both
T4879	7325-7327	IN	denotes	by
T4880	7328-7336	NNP	denotes	Southern
T4881	7337-7341	NN	denotes	blot
T4882	7342-7355	NN	denotes	hybridization
T4883	7356-7359	CC	denotes	and
T4884	7360-7363	NN	denotes	PCR
T4885	7364-7372	NN	denotes	analysis
T4886	7372-7373	.	denotes	.
T4887	7373-7587	sentence	denotes	A probe located outside the short (upstream) arm of homology hybridized to a 3.5-kb BamHI fragment of the wild-type allele and 5.3-kb BamHI fragment of the mutant allele as predicted from genomic maps (Figure 2A).
T4888	7374-7375	DT	denotes	A
T4889	7376-7381	NN	denotes	probe
T4891	7382-7389	VBN	denotes	located
T4892	7390-7397	RB	denotes	outside
T4893	7398-7401	DT	denotes	the
T4895	7402-7407	JJ	denotes	short
T4896	7408-7409	-LRB-	denotes	(
T4897	7409-7417	JJ	denotes	upstream
T4898	7417-7418	-RRB-	denotes	)
T4894	7419-7422	NN	denotes	arm
T4899	7423-7425	IN	denotes	of
T4900	7426-7434	NN	denotes	homology
T4890	7435-7445	VBN	denotes	hybridized
T4901	7446-7448	IN	denotes	to
T4902	7449-7450	DT	denotes	a
T4904	7451-7454	CD	denotes	3.5
T4906	7454-7455	HYPH	denotes	-
T4905	7455-7457	NN	denotes	kb
T4907	7458-7463	NN	denotes	BamHI
T4903	7464-7472	NN	denotes	fragment
T4908	7473-7475	IN	denotes	of
T4909	7476-7479	DT	denotes	the
T4911	7480-7484	JJ	denotes	wild
T4913	7484-7485	HYPH	denotes	-
T4912	7485-7489	NN	denotes	type
T4910	7490-7496	NN	denotes	allele
T4914	7497-7500	CC	denotes	and
T4915	7501-7504	CD	denotes	5.3
T4917	7504-7505	HYPH	denotes	-
T4916	7505-7507	NN	denotes	kb
T4919	7508-7513	NN	denotes	BamHI
T4918	7514-7522	NN	denotes	fragment
T4920	7523-7525	IN	denotes	of
T4921	7526-7529	DT	denotes	the
T4923	7530-7536	NN	denotes	mutant
T4922	7537-7543	NN	denotes	allele
T4924	7544-7546	IN	denotes	as
T4925	7547-7556	VBN	denotes	predicted
T4926	7557-7561	IN	denotes	from
T4927	7562-7569	JJ	denotes	genomic
T4928	7570-7574	NNS	denotes	maps
T4929	7575-7576	-LRB-	denotes	(
T4931	7576-7582	NN	denotes	Figure
T4930	7583-7585	NN	denotes	2A
T4932	7585-7586	-RRB-	denotes	)
T4933	7586-7587	.	denotes	.
T4934	7587-7812	sentence	denotes	The same probe also detected the expected 7.2-kb wild-type and 6.4-kb mutant BglII fragments, 4.9-kb wild-type and 5.7-kb mutant NcoI fragments, as well as 7.2-kb wild-type and 4.9-kb mutant BglII/AgeI fragments (not shown).
T4935	7588-7591	DT	denotes	The
T4937	7592-7596	JJ	denotes	same
T4936	7597-7602	NN	denotes	probe
T4939	7603-7607	RB	denotes	also
T4938	7608-7616	VBD	denotes	detected
T4940	7617-7620	DT	denotes	the
T4942	7621-7629	VBN	denotes	expected
T4943	7630-7633	CD	denotes	7.2
T4945	7633-7634	HYPH	denotes	-
T4944	7634-7636	NN	denotes	kb
T4946	7637-7641	JJ	denotes	wild
T4948	7641-7642	HYPH	denotes	-
T4947	7642-7646	NN	denotes	type
T4949	7647-7650	CC	denotes	and
T4950	7651-7654	CD	denotes	6.4
T4952	7654-7655	HYPH	denotes	-
T4951	7655-7657	NN	denotes	kb
T4953	7658-7664	NN	denotes	mutant
T4954	7665-7670	NN	denotes	BglII
T4941	7671-7680	NNS	denotes	fragments
T4955	7680-7682	,	denotes	,
T4956	7682-7685	CD	denotes	4.9
T4958	7685-7686	HYPH	denotes	-
T4957	7686-7688	NN	denotes	kb
T4960	7689-7693	JJ	denotes	wild
T4962	7693-7694	HYPH	denotes	-
T4961	7694-7698	NN	denotes	type
T4963	7699-7702	CC	denotes	and
T4964	7703-7706	CD	denotes	5.7
T4966	7706-7707	HYPH	denotes	-
T4965	7707-7709	NN	denotes	kb
T4967	7710-7716	NN	denotes	mutant
T4968	7717-7721	NN	denotes	NcoI
T4959	7722-7731	NNS	denotes	fragments
T4969	7731-7733	,	denotes	,
T4970	7733-7735	RB	denotes	as
T4972	7736-7740	RB	denotes	well
T4971	7741-7743	IN	denotes	as
T4973	7744-7747	CD	denotes	7.2
T4975	7747-7748	HYPH	denotes	-
T4974	7748-7750	NN	denotes	kb
T4977	7751-7755	JJ	denotes	wild
T4979	7755-7756	HYPH	denotes	-
T4978	7756-7760	NN	denotes	type
T4980	7761-7764	CC	denotes	and
T4981	7765-7768	CD	denotes	4.9
T4983	7768-7769	HYPH	denotes	-
T4982	7769-7771	NN	denotes	kb
T4984	7772-7778	NN	denotes	mutant
T4985	7779-7784	NN	denotes	BglII
T4987	7784-7785	HYPH	denotes	/
T4986	7785-7789	NN	denotes	AgeI
T4976	7790-7799	NNS	denotes	fragments
T4988	7800-7801	-LRB-	denotes	(
T4990	7801-7804	RB	denotes	not
T4989	7805-7810	VBN	denotes	shown
T4991	7810-7811	-RRB-	denotes	)
T4992	7811-7812	.	denotes	.
T4993	7812-8009	sentence	denotes	A probe derived from the PGK-Neo cassette recognized only the 5.3-kb BamHI fragment in Capn2+/- ES cells, suggesting that the targeting vector had integrated solely at the Capn2 locus (not shown).
T4994	7813-7814	DT	denotes	A
T4995	7815-7820	NN	denotes	probe
T4997	7821-7828	VBN	denotes	derived
T4998	7829-7833	IN	denotes	from
T4999	7834-7837	DT	denotes	the
T5001	7838-7841	NN	denotes	PGK
T5003	7841-7842	HYPH	denotes	-
T5002	7842-7845	NN	denotes	Neo
T5000	7846-7854	NN	denotes	cassette
T4996	7855-7865	VBD	denotes	recognized
T5004	7866-7870	RB	denotes	only
T5006	7871-7874	DT	denotes	the
T5007	7875-7878	CD	denotes	5.3
T5009	7878-7879	HYPH	denotes	-
T5008	7879-7881	NN	denotes	kb
T5010	7882-7887	NN	denotes	BamHI
T5005	7888-7896	NN	denotes	fragment
T5011	7897-7899	IN	denotes	in
T5012	7900-7905	NN	denotes	Capn2
T5014	7905-7906	SYM	denotes	+
T5015	7906-7907	HYPH	denotes	/
T5016	7907-7908	SYM	denotes	-
T5013	7909-7911	NN	denotes	ES
T5017	7912-7917	NNS	denotes	cells
T5018	7917-7919	,	denotes	,
T5019	7919-7929	VBG	denotes	suggesting
T5020	7930-7934	IN	denotes	that
T5022	7935-7938	DT	denotes	the
T5024	7939-7948	NN	denotes	targeting
T5023	7949-7955	NN	denotes	vector
T5025	7956-7959	VBD	denotes	had
T5021	7960-7970	VBN	denotes	integrated
T5026	7971-7977	RB	denotes	solely
T5027	7978-7980	IN	denotes	at
T5028	7981-7984	DT	denotes	the
T5030	7985-7990	NN	denotes	Capn2
T5029	7991-7996	NN	denotes	locus
T5031	7997-7998	-LRB-	denotes	(
T5033	7998-8001	RB	denotes	not
T5032	8002-8007	VBN	denotes	shown
T5034	8007-8008	-RRB-	denotes	)
T5035	8008-8009	.	denotes	.
T5036	8009-8147	sentence	denotes	A PCR screening method was also established that generated a wild-type product of 2,749 bp and a 2,711 bp product from the mutant allele.
T5037	8010-8011	DT	denotes	A
T5039	8012-8015	NN	denotes	PCR
T5040	8016-8025	NN	denotes	screening
T5038	8026-8032	NN	denotes	method
T5042	8033-8036	VBD	denotes	was
T5043	8037-8041	RB	denotes	also
T5041	8042-8053	VBN	denotes	established
T5044	8054-8058	WDT	denotes	that
T5045	8059-8068	VBD	denotes	generated
T5046	8069-8070	DT	denotes	a
T5048	8071-8075	JJ	denotes	wild
T5050	8075-8076	HYPH	denotes	-
T5049	8076-8080	NN	denotes	type
T5047	8081-8088	NN	denotes	product
T5051	8089-8091	IN	denotes	of
T5052	8092-8097	CD	denotes	2,749
T5053	8098-8100	NN	denotes	bp
T5054	8101-8104	CC	denotes	and
T5055	8105-8106	DT	denotes	a
T5057	8107-8112	CD	denotes	2,711
T5058	8113-8115	NN	denotes	bp
T5056	8116-8123	NN	denotes	product
T5059	8124-8128	IN	denotes	from
T5060	8129-8132	DT	denotes	the
T5062	8133-8139	NN	denotes	mutant
T5061	8140-8146	NN	denotes	allele
T5063	8146-8147	.	denotes	.
T5064	8147-8229	sentence	denotes	The 2,711 bp product was only evident in the two targeted cell lines (Figure 2B).
T5065	8148-8151	DT	denotes	The
T5067	8152-8157	CD	denotes	2,711
T5068	8158-8160	NN	denotes	bp
T5066	8161-8168	NN	denotes	product
T5069	8169-8172	VBD	denotes	was
T5070	8173-8177	RB	denotes	only
T5071	8178-8185	JJ	denotes	evident
T5072	8186-8188	IN	denotes	in
T5073	8189-8192	DT	denotes	the
T5075	8193-8196	CD	denotes	two
T5076	8197-8205	VBN	denotes	targeted
T5077	8206-8210	NN	denotes	cell
T5074	8211-8216	NNS	denotes	lines
T5078	8217-8218	-LRB-	denotes	(
T5080	8218-8224	NN	denotes	Figure
T5079	8225-8227	NN	denotes	2B
T5081	8227-8228	-RRB-	denotes	)
T5082	8228-8229	.	denotes	.
T21151	8240-8256	NN	denotes	Characterization
T21152	8257-8259	IN	denotes	of
T21153	8260-8268	VBN	denotes	targeted
T21155	8269-8271	NN	denotes	ES
T21156	8272-8276	NN	denotes	cell
T21154	8277-8282	NNS	denotes	lines
T21157	8282-8283	.	denotes	.
T21158	8283-8371	sentence	denotes	(A) Targeted disruption of the Capn2 locus was detected initially by Southern blotting.
T21159	8284-8285	-LRB-	denotes	(
T21160	8285-8286	LS	denotes	A
T21162	8286-8287	-RRB-	denotes	)
T21163	8288-8296	VBN	denotes	Targeted
T21164	8297-8307	NN	denotes	disruption
T21165	8308-8310	IN	denotes	of
T21166	8311-8314	DT	denotes	the
T21168	8315-8320	NN	denotes	Capn2
T21167	8321-8326	NN	denotes	locus
T21169	8327-8330	VBD	denotes	was
T21161	8331-8339	VBN	denotes	detected
T21170	8340-8349	RB	denotes	initially
T21171	8350-8352	IN	denotes	by
T21172	8353-8361	NNP	denotes	Southern
T21173	8362-8370	NN	denotes	blotting
T21174	8370-8371	.	denotes	.
T21175	8371-8596	sentence	denotes	Membranes were blotted with BamHI-digested genomic DNA extracted from ES cells and hybridized with a DIG-labeled 823 bp BamHI/HindIII fragment located immediately upstream of the short arm of the targeting vector (Figure 1).
T21176	8372-8381	NNS	denotes	Membranes
T21178	8382-8386	VBD	denotes	were
T21177	8387-8394	VBN	denotes	blotted
T21179	8395-8399	IN	denotes	with
T21180	8400-8405	NN	denotes	BamHI
T21182	8405-8406	HYPH	denotes	-
T21181	8406-8414	VBN	denotes	digested
T21184	8415-8422	JJ	denotes	genomic
T21183	8423-8426	NN	denotes	DNA
T21185	8427-8436	VBN	denotes	extracted
T21186	8437-8441	IN	denotes	from
T21187	8442-8444	NN	denotes	ES
T21188	8445-8450	NNS	denotes	cells
T21189	8451-8454	CC	denotes	and
T21190	8455-8465	VBD	denotes	hybridized
T21191	8466-8470	IN	denotes	with
T21192	8471-8472	DT	denotes	a
T21194	8473-8476	NN	denotes	DIG
T21196	8476-8477	HYPH	denotes	-
T21195	8477-8484	VBN	denotes	labeled
T21197	8485-8488	CD	denotes	823
T21198	8489-8491	NN	denotes	bp
T21199	8492-8497	NN	denotes	BamHI
T21201	8497-8498	HYPH	denotes	/
T21200	8498-8505	NN	denotes	HindIII
T21193	8506-8514	NN	denotes	fragment
T21202	8515-8522	VBN	denotes	located
T21203	8523-8534	RB	denotes	immediately
T21204	8535-8543	IN	denotes	upstream
T21205	8544-8546	IN	denotes	of
T21206	8547-8550	DT	denotes	the
T21208	8551-8556	JJ	denotes	short
T21207	8557-8560	NN	denotes	arm
T21209	8561-8563	IN	denotes	of
T21210	8564-8567	DT	denotes	the
T21212	8568-8577	NN	denotes	targeting
T21211	8578-8584	NN	denotes	vector
T21213	8585-8586	-LRB-	denotes	(
T21214	8586-8592	NN	denotes	Figure
T21215	8593-8594	CD	denotes	1
T21216	8594-8595	-RRB-	denotes	)
T21217	8595-8596	.	denotes	.
T21218	8596-8800	sentence	denotes	A 3.5-kb BamHI fragment corresponding to the wild-type allele was present in all cells, whereas a 5.3-kb fragment from the mutant allele was detected in two targeted cell lines, designated ES27 and ES36.
T21219	8597-8598	DT	denotes	A
T21221	8599-8602	CD	denotes	3.5
T21223	8602-8603	HYPH	denotes	-
T21222	8603-8605	NN	denotes	kb
T21224	8606-8611	NN	denotes	BamHI
T21220	8612-8620	NN	denotes	fragment
T21226	8621-8634	VBG	denotes	corresponding
T21227	8635-8637	IN	denotes	to
T21228	8638-8641	DT	denotes	the
T21230	8642-8646	JJ	denotes	wild
T21232	8646-8647	HYPH	denotes	-
T21231	8647-8651	NN	denotes	type
T21229	8652-8658	NN	denotes	allele
T21225	8659-8662	VBD	denotes	was
T21233	8663-8670	JJ	denotes	present
T21234	8671-8673	IN	denotes	in
T21235	8674-8677	DT	denotes	all
T21236	8678-8683	NNS	denotes	cells
T21237	8683-8685	,	denotes	,
T21238	8685-8692	IN	denotes	whereas
T21240	8693-8694	DT	denotes	a
T21242	8695-8698	CD	denotes	5.3
T21244	8698-8699	HYPH	denotes	-
T21243	8699-8701	NN	denotes	kb
T21241	8702-8710	NN	denotes	fragment
T21245	8711-8715	IN	denotes	from
T21246	8716-8719	DT	denotes	the
T21248	8720-8726	NN	denotes	mutant
T21247	8727-8733	NN	denotes	allele
T21249	8734-8737	VBD	denotes	was
T21239	8738-8746	VBN	denotes	detected
T21250	8747-8749	IN	denotes	in
T21251	8750-8753	CD	denotes	two
T21253	8754-8762	VBN	denotes	targeted
T21254	8763-8767	NN	denotes	cell
T21252	8768-8773	NNS	denotes	lines
T21255	8773-8775	,	denotes	,
T21256	8775-8785	VBN	denotes	designated
T21257	8786-8790	NN	denotes	ES27
T21258	8791-8794	CC	denotes	and
T21259	8795-8799	NN	denotes	ES36
T21260	8799-8800	.	denotes	.
T21261	8800-8981	sentence	denotes	(B) PCR genotyping was carried out with two separate reactions designed to amplify either a 2,748 bp segment from the wild-type allele or a 2,711 bp segment from the mutant allele.
T21262	8801-8802	-LRB-	denotes	(
T21263	8802-8803	LS	denotes	B
T21265	8803-8804	-RRB-	denotes	)
T21266	8805-8808	NN	denotes	PCR
T21267	8809-8819	NN	denotes	genotyping
T21268	8820-8823	VBD	denotes	was
T21264	8824-8831	VBN	denotes	carried
T21269	8832-8835	RP	denotes	out
T21270	8836-8840	IN	denotes	with
T21271	8841-8844	CD	denotes	two
T21273	8845-8853	JJ	denotes	separate
T21272	8854-8863	NNS	denotes	reactions
T21274	8864-8872	VBN	denotes	designed
T21275	8873-8875	TO	denotes	to
T21276	8876-8883	VB	denotes	amplify
T21277	8884-8890	CC	denotes	either
T21279	8891-8892	DT	denotes	a
T21280	8893-8898	CD	denotes	2,748
T21281	8899-8901	NN	denotes	bp
T21278	8902-8909	NN	denotes	segment
T21282	8910-8914	IN	denotes	from
T21283	8915-8918	DT	denotes	the
T21285	8919-8923	JJ	denotes	wild
T21287	8923-8924	HYPH	denotes	-
T21286	8924-8928	NN	denotes	type
T21284	8929-8935	NN	denotes	allele
T21288	8936-8938	CC	denotes	or
T21289	8939-8940	DT	denotes	a
T21291	8941-8946	CD	denotes	2,711
T21292	8947-8949	NN	denotes	bp
T21290	8950-8957	NN	denotes	segment
T21293	8958-8962	IN	denotes	from
T21294	8963-8966	DT	denotes	the
T21296	8967-8973	NN	denotes	mutant
T21295	8974-8980	NN	denotes	allele
T21297	8980-8981	.	denotes	.
T21298	8981-9139	sentence	denotes	Both reactions used a common sense primer located in intron 4, outside the short arm of the targeting vector, and distinct allele-specific antisense primers.
T21299	8982-8986	DT	denotes	Both
T21300	8987-8996	NNS	denotes	reactions
T21301	8997-9001	VBD	denotes	used
T21302	9002-9003	DT	denotes	a
T21304	9004-9010	JJ	denotes	common
T21305	9011-9016	NN	denotes	sense
T21303	9017-9023	NN	denotes	primer
T21306	9024-9031	VBN	denotes	located
T21307	9032-9034	IN	denotes	in
T21308	9035-9041	NN	denotes	intron
T21309	9042-9043	CD	denotes	4
T21310	9043-9045	,	denotes	,
T21311	9045-9052	IN	denotes	outside
T21312	9053-9056	DT	denotes	the
T21314	9057-9062	JJ	denotes	short
T21313	9063-9066	NN	denotes	arm
T21315	9067-9069	IN	denotes	of
T21316	9070-9073	DT	denotes	the
T21318	9074-9083	NN	denotes	targeting
T21317	9084-9090	NN	denotes	vector
T21319	9090-9092	,	denotes	,
T21320	9092-9095	CC	denotes	and
T21321	9096-9104	JJ	denotes	distinct
T21323	9105-9111	NN	denotes	allele
T21325	9111-9112	HYPH	denotes	-
T21324	9112-9120	JJ	denotes	specific
T21326	9121-9130	NN	denotes	antisense
T21322	9131-9138	NNS	denotes	primers
T21327	9138-9139	.	denotes	.
T21328	9139-9332	sentence	denotes	The reaction to detect the wild-type allele used an antisense primer located in exon 7 while the amplification of the mutant sequence was done with an antisense primer in the PGK-Neo cassette.
T21329	9140-9143	DT	denotes	The
T21330	9144-9152	NN	denotes	reaction
T21332	9153-9155	TO	denotes	to
T21333	9156-9162	VB	denotes	detect
T21334	9163-9166	DT	denotes	the
T21336	9167-9171	JJ	denotes	wild
T21338	9171-9172	HYPH	denotes	-
T21337	9172-9176	NN	denotes	type
T21335	9177-9183	NN	denotes	allele
T21331	9184-9188	VBD	denotes	used
T21339	9189-9191	DT	denotes	an
T21341	9192-9201	NN	denotes	antisense
T21340	9202-9208	NN	denotes	primer
T21342	9209-9216	VBN	denotes	located
T21343	9217-9219	IN	denotes	in
T21344	9220-9224	NN	denotes	exon
T21345	9225-9226	CD	denotes	7
T21346	9227-9232	IN	denotes	while
T21348	9233-9236	DT	denotes	the
T21349	9237-9250	NN	denotes	amplification
T21350	9251-9253	IN	denotes	of
T21351	9254-9257	DT	denotes	the
T21353	9258-9264	NN	denotes	mutant
T21352	9265-9273	NN	denotes	sequence
T21354	9274-9277	VBD	denotes	was
T21347	9278-9282	VBN	denotes	done
T21355	9283-9287	IN	denotes	with
T21356	9288-9290	DT	denotes	an
T21358	9291-9300	NN	denotes	antisense
T21357	9301-9307	NN	denotes	primer
T21359	9308-9310	IN	denotes	in
T21360	9311-9314	DT	denotes	the
T21362	9315-9318	NN	denotes	PGK
T21364	9318-9319	HYPH	denotes	-
T21363	9319-9322	NN	denotes	Neo
T21361	9323-9331	NN	denotes	cassette
T21365	9331-9332	.	denotes	.
T21366	9332-9482	sentence	denotes	The results confirm the presence of the wild-type allele in all cells, whereas the mutant allele signal was observed only in the two targeted clones.
T21367	9333-9336	DT	denotes	The
T21368	9337-9344	NNS	denotes	results
T21369	9345-9352	VBP	denotes	confirm
T21370	9353-9356	DT	denotes	the
T21371	9357-9365	NN	denotes	presence
T21372	9366-9368	IN	denotes	of
T21373	9369-9372	DT	denotes	the
T21375	9373-9377	JJ	denotes	wild
T21377	9377-9378	HYPH	denotes	-
T21376	9378-9382	NN	denotes	type
T21374	9383-9389	NN	denotes	allele
T21378	9390-9392	IN	denotes	in
T21379	9393-9396	DT	denotes	all
T21380	9397-9402	NNS	denotes	cells
T21381	9402-9404	,	denotes	,
T21382	9404-9411	IN	denotes	whereas
T21384	9412-9415	DT	denotes	the
T21386	9416-9422	NN	denotes	mutant
T21387	9423-9429	NN	denotes	allele
T21385	9430-9436	NN	denotes	signal
T21388	9437-9440	VBD	denotes	was
T21383	9441-9449	VBN	denotes	observed
T21389	9450-9454	RB	denotes	only
T21390	9455-9457	IN	denotes	in
T21391	9458-9461	DT	denotes	the
T21393	9462-9465	CD	denotes	two
T21394	9466-9474	VBN	denotes	targeted
T21392	9475-9481	NNS	denotes	clones
T21395	9481-9482	.	denotes	.
T21396	9482-9523	sentence	denotes	(M) denotes the molecular weight marker.
T21397	9483-9484	-LRB-	denotes	(
T21399	9484-9485	NN	denotes	M
T21400	9485-9486	-RRB-	denotes	)
T21398	9487-9494	VBZ	denotes	denotes
T21401	9495-9498	DT	denotes	the
T21403	9499-9508	JJ	denotes	molecular
T21404	9509-9515	NN	denotes	weight
T21402	9516-9522	NN	denotes	marker
T21405	9522-9523	.	denotes	.
T6635	9525-9535	NN	denotes	Generation
T6636	9536-9538	IN	denotes	of
T6637	9539-9547	JJ	denotes	chimeric
T6638	9548-9552	NNS	denotes	mice
T6639	9553-9556	CC	denotes	and
T6640	9557-9565	NN	denotes	germline
T6641	9566-9578	NN	denotes	transmission
T6642	9579-9581	IN	denotes	of
T6643	9582-9585	DT	denotes	the
T6645	9586-9592	NN	denotes	mutant
T6646	9593-9598	NN	denotes	Capn2
T6644	9599-9605	NN	denotes	allele
T6647	9605-9713	sentence	denotes	Eight chimeric male mice were produced in morula aggregation experiments using the Capn2+/- ES27 cell line.
T6648	9606-9611	CD	denotes	Eight
T6650	9612-9620	JJ	denotes	chimeric
T6651	9621-9625	JJ	denotes	male
T6649	9626-9630	NNS	denotes	mice
T6653	9631-9635	VBD	denotes	were
T6652	9636-9644	VBN	denotes	produced
T6654	9645-9647	IN	denotes	in
T6655	9648-9654	NN	denotes	morula
T6656	9655-9666	NN	denotes	aggregation
T6657	9667-9678	NNS	denotes	experiments
T6658	9679-9684	VBG	denotes	using
T6659	9685-9688	DT	denotes	the
T6661	9689-9694	NN	denotes	Capn2
T6662	9694-9695	SYM	denotes	+
T6663	9695-9696	HYPH	denotes	/
T6664	9696-9697	SYM	denotes	-
T6665	9698-9702	NN	denotes	ES27
T6666	9703-9707	NN	denotes	cell
T6660	9708-9712	NN	denotes	line
T6667	9712-9713	.	denotes	.
T6668	9713-9816	sentence	denotes	Two of these males transmitted the Capn2+/- ES27 genotype through the germline into the F1 generation.
T6669	9714-9717	CD	denotes	Two
T6671	9718-9720	IN	denotes	of
T6672	9721-9726	DT	denotes	these
T6673	9727-9732	NNS	denotes	males
T6670	9733-9744	VBD	denotes	transmitted
T6674	9745-9748	DT	denotes	the
T6676	9749-9754	NN	denotes	Capn2
T6677	9754-9755	SYM	denotes	+
T6678	9755-9756	HYPH	denotes	/
T6679	9756-9757	SYM	denotes	-
T6680	9758-9762	NN	denotes	ES27
T6675	9763-9771	NN	denotes	genotype
T6681	9772-9779	IN	denotes	through
T6682	9780-9783	DT	denotes	the
T6683	9784-9792	NN	denotes	germline
T6684	9793-9797	IN	denotes	into
T6685	9798-9801	DT	denotes	the
T6687	9802-9804	NN	denotes	F1
T6686	9805-9815	NN	denotes	generation
T6688	9815-9816	.	denotes	.
T6689	9816-9933	sentence	denotes	Heterozygous Capn2+/- animals appeared normal, with no obvious defects in gross anatomy, reproduction, or life span.
T6690	9817-9829	JJ	denotes	Heterozygous
T6692	9830-9835	NN	denotes	Capn2
T6693	9835-9836	SYM	denotes	+
T6694	9836-9837	HYPH	denotes	/
T6695	9837-9838	SYM	denotes	-
T6691	9839-9846	NNS	denotes	animals
T6696	9847-9855	VBD	denotes	appeared
T6697	9856-9862	JJ	denotes	normal
T6698	9862-9864	,	denotes	,
T6699	9864-9868	IN	denotes	with
T6700	9869-9871	DT	denotes	no
T6702	9872-9879	JJ	denotes	obvious
T6701	9880-9887	NNS	denotes	defects
T6703	9888-9890	IN	denotes	in
T6704	9891-9896	JJ	denotes	gross
T6705	9897-9904	NN	denotes	anatomy
T6706	9904-9906	,	denotes	,
T6707	9906-9918	NN	denotes	reproduction
T6708	9918-9920	,	denotes	,
T6709	9920-9922	CC	denotes	or
T6710	9923-9927	NN	denotes	life
T6711	9928-9932	NN	denotes	span
T6712	9932-9933	.	denotes	.
T6713	9933-10031	sentence	denotes	Out of 199 weanlings from heterozygous intercrosses, no Capn2-/- progeny were detected (Table 2).
T6714	9934-9937	IN	denotes	Out
T6716	9938-9940	IN	denotes	of
T6717	9941-9944	CD	denotes	199
T6718	9945-9954	NNS	denotes	weanlings
T6719	9955-9959	IN	denotes	from
T6720	9960-9972	JJ	denotes	heterozygous
T6721	9973-9985	NNS	denotes	intercrosses
T6722	9985-9987	,	denotes	,
T6723	9987-9989	DT	denotes	no
T6725	9990-9995	NN	denotes	Capn2
T6726	9995-9996	SYM	denotes	-
T6727	9996-9997	HYPH	denotes	/
T6728	9997-9998	SYM	denotes	-
T6724	9999-10006	NN	denotes	progeny
T6729	10007-10011	VBD	denotes	were
T6715	10012-10020	VBN	denotes	detected
T6730	10021-10022	-LRB-	denotes	(
T6731	10022-10027	NN	denotes	Table
T6732	10028-10029	CD	denotes	2
T6733	10029-10030	-RRB-	denotes	)
T6734	10030-10031	.	denotes	.
T6735	10031-10123	sentence	denotes	We did not observe high rates of perinatal death, and no Capn2-/- stillborns were observed.
T6736	10032-10034	PRP	denotes	We
T6738	10035-10038	VBD	denotes	did
T6739	10039-10042	RB	denotes	not
T6737	10043-10050	VB	denotes	observe
T6740	10051-10055	JJ	denotes	high
T6741	10056-10061	NNS	denotes	rates
T6742	10062-10064	IN	denotes	of
T6743	10065-10074	JJ	denotes	perinatal
T6744	10075-10080	NN	denotes	death
T6745	10080-10082	,	denotes	,
T6746	10082-10085	CC	denotes	and
T6747	10086-10088	DT	denotes	no
T6749	10089-10094	NN	denotes	Capn2
T6750	10094-10095	SYM	denotes	-
T6751	10095-10096	HYPH	denotes	/
T6752	10096-10097	SYM	denotes	-
T6748	10098-10108	NNS	denotes	stillborns
T6754	10109-10113	VBD	denotes	were
T6753	10114-10122	VBN	denotes	observed
T6755	10122-10123	.	denotes	.
T6756	10123-10213	sentence	denotes	This suggested that Capn2-/- animals perished at some stage during embryonic development.
T6757	10124-10128	DT	denotes	This
T6758	10129-10138	VBD	denotes	suggested
T6759	10139-10143	IN	denotes	that
T6761	10144-10149	NN	denotes	Capn2
T6763	10149-10150	SYM	denotes	-
T6764	10150-10151	HYPH	denotes	/
T6765	10151-10152	SYM	denotes	-
T6762	10153-10160	NNS	denotes	animals
T6760	10161-10169	VBN	denotes	perished
T6766	10170-10172	IN	denotes	at
T6767	10173-10177	DT	denotes	some
T6768	10178-10183	NN	denotes	stage
T6769	10184-10190	IN	denotes	during
T6770	10191-10200	JJ	denotes	embryonic
T6771	10201-10212	NN	denotes	development
T6772	10212-10213	.	denotes	.
T6773	10213-10380	sentence	denotes	In an attempt to determine if embryonic death occurred at a post-implantation stage, embryos were harvested for genotyping at different times between E10.5 and E18.5.
T6774	10214-10216	IN	denotes	In
T6776	10217-10219	DT	denotes	an
T6777	10220-10227	NN	denotes	attempt
T6778	10228-10230	TO	denotes	to
T6779	10231-10240	VB	denotes	determine
T6780	10241-10243	IN	denotes	if
T6782	10244-10253	JJ	denotes	embryonic
T6783	10254-10259	NN	denotes	death
T6781	10260-10268	VBD	denotes	occurred
T6784	10269-10271	IN	denotes	at
T6785	10272-10273	DT	denotes	a
T6787	10274-10291	JJ	denotes	post-implantation
T6786	10292-10297	NN	denotes	stage
T6788	10297-10299	,	denotes	,
T6789	10299-10306	NNS	denotes	embryos
T6790	10307-10311	VBD	denotes	were
T6775	10312-10321	VBN	denotes	harvested
T6791	10322-10325	IN	denotes	for
T6792	10326-10336	NN	denotes	genotyping
T6793	10337-10339	IN	denotes	at
T6794	10340-10349	JJ	denotes	different
T6795	10350-10355	NNS	denotes	times
T6796	10356-10363	IN	denotes	between
T6797	10364-10369	NN	denotes	E10.5
T6798	10370-10373	CC	denotes	and
T6799	10374-10379	NN	denotes	E18.5
T6800	10379-10380	.	denotes	.
T6801	10380-10473	sentence	denotes	No Capn2-/- embryos were observed and no signs of embryo resorption were detected (Table 2).
T6802	10381-10383	DT	denotes	No
T6804	10384-10389	NN	denotes	Capn2
T6805	10389-10390	SYM	denotes	-
T6806	10390-10391	HYPH	denotes	/
T6807	10391-10392	SYM	denotes	-
T6803	10393-10400	NNS	denotes	embryos
T6809	10401-10405	VBD	denotes	were
T6808	10406-10414	VBN	denotes	observed
T6810	10415-10418	CC	denotes	and
T6811	10419-10421	DT	denotes	no
T6812	10422-10427	NNS	denotes	signs
T6814	10428-10430	IN	denotes	of
T6815	10431-10437	NN	denotes	embryo
T6816	10438-10448	NN	denotes	resorption
T6817	10449-10453	VBD	denotes	were
T6813	10454-10462	VBN	denotes	detected
T6818	10463-10464	-LRB-	denotes	(
T6819	10464-10469	NN	denotes	Table
T6820	10470-10471	CD	denotes	2
T6821	10471-10472	-RRB-	denotes	)
T6822	10472-10473	.	denotes	.
T6823	10473-10552	sentence	denotes	This indicated that the Capn2-/- embryos might be dying prior to implantation.
T6824	10474-10478	DT	denotes	This
T6825	10479-10488	VBD	denotes	indicated
T6826	10489-10493	IN	denotes	that
T6828	10494-10497	DT	denotes	the
T6830	10498-10503	NN	denotes	Capn2
T6831	10503-10504	SYM	denotes	-
T6832	10504-10505	HYPH	denotes	/
T6833	10505-10506	SYM	denotes	-
T6829	10507-10514	NNS	denotes	embryos
T6834	10515-10520	MD	denotes	might
T6835	10521-10523	VB	denotes	be
T6827	10524-10529	VBG	denotes	dying
T6836	10530-10535	JJ	denotes	prior
T6837	10536-10538	IN	denotes	to
T6838	10539-10551	NN	denotes	implantation
T6839	10551-10552	.	denotes	.
T6840	10552-10693	sentence	denotes	Embryos were then flushed from the oviducts of pregnant females at E2.5 or E3.5, and genotyped by means of a nested PCR strategy (Figure 4).
T6841	10553-10560	NNS	denotes	Embryos
T6843	10561-10565	VBD	denotes	were
T6844	10566-10570	RB	denotes	then
T6842	10571-10578	VBN	denotes	flushed
T6845	10579-10583	IN	denotes	from
T6846	10584-10587	DT	denotes	the
T6847	10588-10596	NNS	denotes	oviducts
T6848	10597-10599	IN	denotes	of
T6849	10600-10608	JJ	denotes	pregnant
T6850	10609-10616	NNS	denotes	females
T6851	10617-10619	IN	denotes	at
T6852	10620-10624	NN	denotes	E2.5
T6853	10625-10627	CC	denotes	or
T6854	10628-10632	NN	denotes	E3.5
T6855	10632-10634	,	denotes	,
T6856	10634-10637	CC	denotes	and
T6857	10638-10647	VBN	denotes	genotyped
T6858	10648-10650	IN	denotes	by
T6859	10651-10656	NNS	denotes	means
T6860	10657-10659	IN	denotes	of
T6861	10660-10661	DT	denotes	a
T6863	10662-10668	VBN	denotes	nested
T6864	10669-10672	NN	denotes	PCR
T6862	10673-10681	NN	denotes	strategy
T6865	10682-10683	-LRB-	denotes	(
T6866	10683-10689	NN	denotes	Figure
T6867	10690-10691	CD	denotes	4
T6868	10691-10692	-RRB-	denotes	)
T6869	10692-10693	.	denotes	.
T6870	10693-10787	sentence	denotes	Two of 90 successfully genotyped pre-implantation embryos were Capn2-/-, (Table 2; Figure 5).
T6871	10694-10697	CD	denotes	Two
T6873	10698-10700	IN	denotes	of
T6874	10701-10703	CD	denotes	90
T6876	10704-10716	RB	denotes	successfully
T6877	10717-10726	VBN	denotes	genotyped
T6878	10727-10743	JJ	denotes	pre-implantation
T6875	10744-10751	NNS	denotes	embryos
T6872	10752-10756	VBD	denotes	were
T6879	10757-10762	NN	denotes	Capn2
T6880	10762-10763	SYM	denotes	-
T6881	10763-10764	HYPH	denotes	/
T6882	10764-10765	SYM	denotes	-
T6883	10765-10767	,	denotes	,
T6884	10767-10768	-LRB-	denotes	(
T6886	10768-10773	NN	denotes	Table
T6887	10774-10775	CD	denotes	2
T6888	10775-10776	:	denotes	;
T6885	10777-10783	NN	denotes	Figure
T6889	10784-10785	CD	denotes	5
T6890	10785-10786	-RRB-	denotes	)
T6891	10786-10787	.	denotes	.
T6892	10787-10907	sentence	denotes	Both of these Capn2-/- embryos were isolated at the 8-cell stage and did not display any obvious morphological defects.
T6893	10788-10792	DT	denotes	Both
T6895	10793-10795	IN	denotes	of
T6896	10796-10801	DT	denotes	these
T6898	10802-10807	NN	denotes	Capn2
T6899	10807-10808	SYM	denotes	-
T6900	10808-10809	HYPH	denotes	/
T6901	10809-10810	SYM	denotes	-
T6897	10811-10818	NNS	denotes	embryos
T6902	10819-10823	VBD	denotes	were
T6894	10824-10832	VBN	denotes	isolated
T6903	10833-10835	IN	denotes	at
T6904	10836-10839	DT	denotes	the
T6906	10840-10841	CD	denotes	8
T6908	10841-10842	HYPH	denotes	-
T6907	10842-10846	NN	denotes	cell
T6905	10847-10852	NN	denotes	stage
T6909	10853-10856	CC	denotes	and
T6910	10857-10860	VBD	denotes	did
T6912	10861-10864	RB	denotes	not
T6911	10865-10872	VB	denotes	display
T6913	10873-10876	DT	denotes	any
T6915	10877-10884	JJ	denotes	obvious
T6916	10885-10898	JJ	denotes	morphological
T6914	10899-10906	NNS	denotes	defects
T6917	10906-10907	.	denotes	.
T6918	10907-10986	sentence	denotes	None of the 46 successfully genotyped blastocyst-staged embryos were Capn2-/-.
T6919	10908-10912	NN	denotes	None
T6921	10913-10915	IN	denotes	of
T6922	10916-10919	DT	denotes	the
T6924	10920-10922	CD	denotes	46
T6925	10923-10935	RB	denotes	successfully
T6926	10936-10945	VBN	denotes	genotyped
T6927	10946-10956	NN	denotes	blastocyst
T6929	10956-10957	HYPH	denotes	-
T6928	10957-10963	VBN	denotes	staged
T6923	10964-10971	NNS	denotes	embryos
T6920	10972-10976	VBD	denotes	were
T6930	10977-10982	NN	denotes	Capn2
T6931	10982-10983	SYM	denotes	-
T6932	10983-10984	HYPH	denotes	/
T6933	10984-10985	SYM	denotes	-
T6934	10985-10986	.	denotes	.
T6935	10986-11158	sentence	denotes	The scarcity of Capn2-deficient embryos surviving to the 8-cell stage suggested that the loss of m-calpain activity must fatally compromise the viability of early embryos.
T6936	10987-10990	DT	denotes	The
T6937	10991-10999	NN	denotes	scarcity
T6939	11000-11002	IN	denotes	of
T6940	11003-11008	NN	denotes	Capn2
T6942	11008-11009	HYPH	denotes	-
T6941	11009-11018	JJ	denotes	deficient
T6943	11019-11026	NNS	denotes	embryos
T6944	11027-11036	VBG	denotes	surviving
T6945	11037-11039	IN	denotes	to
T6946	11040-11043	DT	denotes	the
T6948	11044-11045	CD	denotes	8
T6950	11045-11046	HYPH	denotes	-
T6949	11046-11050	NN	denotes	cell
T6947	11051-11056	NN	denotes	stage
T6938	11057-11066	VBD	denotes	suggested
T6951	11067-11071	IN	denotes	that
T6953	11072-11075	DT	denotes	the
T6954	11076-11080	NN	denotes	loss
T6955	11081-11083	IN	denotes	of
T6956	11084-11085	NN	denotes	m
T6958	11085-11086	HYPH	denotes	-
T6957	11086-11093	NN	denotes	calpain
T6959	11094-11102	NN	denotes	activity
T6960	11103-11107	MD	denotes	must
T6961	11108-11115	RB	denotes	fatally
T6952	11116-11126	VB	denotes	compromise
T6962	11127-11130	DT	denotes	the
T6963	11131-11140	NN	denotes	viability
T6964	11141-11143	IN	denotes	of
T6965	11144-11149	JJ	denotes	early
T6966	11150-11157	NNS	denotes	embryos
T6967	11157-11158	.	denotes	.
T6968	11158-11343	sentence	denotes	Furthermore, it is possible that persistence of some maternally derived mRNA transcript or protein might have allowed a small number of Capn2-/- embryos to survive to the morula-stage.
T6969	11159-11170	RB	denotes	Furthermore
T6971	11170-11172	,	denotes	,
T6972	11172-11174	PRP	denotes	it
T6970	11175-11177	VBZ	denotes	is
T6973	11178-11186	JJ	denotes	possible
T6974	11187-11191	IN	denotes	that
T6976	11192-11203	NN	denotes	persistence
T6977	11204-11206	IN	denotes	of
T6978	11207-11211	DT	denotes	some
T6980	11212-11222	RB	denotes	maternally
T6981	11223-11230	VBN	denotes	derived
T6982	11231-11235	NN	denotes	mRNA
T6979	11236-11246	NN	denotes	transcript
T6983	11247-11249	CC	denotes	or
T6984	11250-11257	NN	denotes	protein
T6985	11258-11263	MD	denotes	might
T6986	11264-11268	VB	denotes	have
T6975	11269-11276	VBN	denotes	allowed
T6987	11277-11278	DT	denotes	a
T6989	11279-11284	JJ	denotes	small
T6988	11285-11291	NN	denotes	number
T6991	11292-11294	IN	denotes	of
T6992	11295-11300	NN	denotes	Capn2
T6994	11300-11301	SYM	denotes	-
T6995	11301-11302	HYPH	denotes	/
T6996	11302-11303	SYM	denotes	-
T6993	11304-11311	NNS	denotes	embryos
T6997	11312-11314	TO	denotes	to
T6990	11315-11322	VB	denotes	survive
T6998	11323-11325	IN	denotes	to
T6999	11326-11329	DT	denotes	the
T7001	11330-11336	NN	denotes	morula
T7002	11336-11337	HYPH	denotes	-
T7000	11337-11342	NN	denotes	stage
T7003	11342-11343	.	denotes	.
T7004	11343-14315	sentence	denotes	Table 2 Genotype distribution of offspring derived from Capn2 transgenic mice. Cross (N) Age Genotype +/+ +/- -/- ND* (+/-)♀ × (+/-)♂ 199 3 weeks 23 176 0 0 (+/+)♀ × (+/-)♂ 122 3 weeks 50 72 0 0 (+/-)♀ × (+/+)♂ 103 3 weeks 28 75 0 0 (+/-)♀ × (+/-)♂ 9 E18.5 0 7 0 2 (+/-)♀ × (+/-)♂ 5 E17.5 0 5 0 0 (+/-)♀ × (+/-)♂ 9 E14.5 2 7 0 0 (+/-)♀ × (+/-)♂ 10 E11.5 3 7 0 0 (+/-)♀ × (+/-)♂ 7 E10.5 2 5 0 0 (+/-)♀ × (+/-)♂ 46 E3.5 7 39 0 0 (+/-)♀ × (+/-)♂ 48 E2.5 8 34 2 4 * ND, not determined Figure 4 Nested PCR strategy for genotyping of pre-implantation embryos. Due to the scarcity of extractable genetic material, a nested PCR strategy was developed in order to genotype pre-implantation embryos. Separate reactions were used to amplify a 429 bp fragment from the wild-type allele and a 389 bp segment from the mutant allele, both spanning the 3' end of the short (upstream) arm of the targeting vector. In both reactions, a 213 bp sequence located within the short arm was co-amplified with the 'diagnostic' products as an internal control. The same sense primers were used to amplify 'diagnostic' sequences in both reactions, whereas the antisense primers were allele-specific. The primers, represented by triangles, are depicted in two (nested) sets for each of the three reactions. Exons are represented by open, vertical rectangles, the PGK-Neo cassette by an open, horizontal rectangle, while thin vertical lines denote the boundaries of the short arm and the 5' end of the long (downstream) arm. A grey, horizontal rectangle delineates the segment of the wild-type allele that is replaced by the PGK-Neo cassette in the mutant allele. Figure 5 Genotyping of pre-implantation embryos. A nested PCR strategy was used to genotype embryos prior to implantation as described in Figure 4. Capn2+/- mice were mated and the date of fertilization established by the appearance of vaginal plugs. Blastocyst (E3.5) or 8-cell embryos (E2.5) were flushed from the oviducts and then digested with proteinase K. In separate reactions segments found exclusively in either the wild-type or mutant alleles were co-amplified with an internal control sequence, located in the short (upstream) arm of the targeting vector, which is found in both alleles. The final products were 429 bp for the wild-type allele, 389 bp for the mutant allele, and 213 bp for the internal control. (A) A representative example of the genotyping of blastocyst stage embryos. Embryos #1, 2, 4, 5, and 6 were Capn2+/- whereas embryos #3 and #7 were Capn2+/+, denoted by the absence of the 389 bp mutant signal. (B) An example of the genotyping of 8-cell embryos is shown. Embryos #1, 2, 3, 5, and 6 were Capn2+/- while embryo #4 was Capn2-/-, marked by the absence of the 429 bp wild-type signal. (M) denotes the molecular weight marker. The genotyping results for weanlings, post-implantation, and pre-implantation embryos are shown in Table 2.
T7005	11344-14211	DT	denotes	Table 2 Genotype distribution of offspring derived from Capn2 transgenic mice. Cross (N) Age Genotype +/+ +/- -/- ND* (+/-)♀ × (+/-)♂ 199 3 weeks 23 176 0 0 (+/+)♀ × (+/-)♂ 122 3 weeks 50 72 0 0 (+/-)♀ × (+/+)♂ 103 3 weeks 28 75 0 0 (+/-)♀ × (+/-)♂ 9 E18.5 0 7 0 2 (+/-)♀ × (+/-)♂ 5 E17.5 0 5 0 0 (+/-)♀ × (+/-)♂ 9 E14.5 2 7 0 0 (+/-)♀ × (+/-)♂ 10 E11.5 3 7 0 0 (+/-)♀ × (+/-)♂ 7 E10.5 2 5 0 0 (+/-)♀ × (+/-)♂ 46 E3.5 7 39 0 0 (+/-)♀ × (+/-)♂ 48 E2.5 8 34 2 4 * ND, not determined Figure 4 Nested PCR strategy for genotyping of pre-implantation embryos. Due to the scarcity of extractable genetic material, a nested PCR strategy was developed in order to genotype pre-implantation embryos. Separate reactions were used to amplify a 429 bp fragment from the wild-type allele and a 389 bp segment from the mutant allele, both spanning the 3' end of the short (upstream) arm of the targeting vector. In both reactions, a 213 bp sequence located within the short arm was co-amplified with the 'diagnostic' products as an internal control. The same sense primers were used to amplify 'diagnostic' sequences in both reactions, whereas the antisense primers were allele-specific. The primers, represented by triangles, are depicted in two (nested) sets for each of the three reactions. Exons are represented by open, vertical rectangles, the PGK-Neo cassette by an open, horizontal rectangle, while thin vertical lines denote the boundaries of the short arm and the 5' end of the long (downstream) arm. A grey, horizontal rectangle delineates the segment of the wild-type allele that is replaced by the PGK-Neo cassette in the mutant allele. Figure 5 Genotyping of pre-implantation embryos. A nested PCR strategy was used to genotype embryos prior to implantation as described in Figure 4. Capn2+/- mice were mated and the date of fertilization established by the appearance of vaginal plugs. Blastocyst (E3.5) or 8-cell embryos (E2.5) were flushed from the oviducts and then digested with proteinase K. In separate reactions segments found exclusively in either the wild-type or mutant alleles were co-amplified with an internal control sequence, located in the short (upstream) arm of the targeting vector, which is found in both alleles. The final products were 429 bp for the wild-type allele, 389 bp for the mutant allele, and 213 bp for the internal control. (A) A representative example of the genotyping of blastocyst stage embryos. Embryos #1, 2, 4, 5, and 6 were Capn2+/- whereas embryos #3 and #7 were Capn2+/+, denoted by the absence of the 389 bp mutant signal. (B) An example of the genotyping of 8-cell embryos is shown. Embryos #1, 2, 3, 5, and 6 were Capn2+/- while embryo #4 was Capn2-/-, marked by the absence of the 429 bp wild-type signal. (M) denotes the molecular weight marker. The
T22936	11353-11361	NN	denotes	Genotype
T22937	11362-11374	NN	denotes	distribution
T22938	11375-11377	IN	denotes	of
T22939	11378-11387	NN	denotes	offspring
T22940	11388-11395	VBN	denotes	derived
T22941	11396-11400	IN	denotes	from
T22942	11401-11406	NN	denotes	Capn2
T22944	11407-11417	JJ	denotes	transgenic
T22943	11418-11422	NNS	denotes	mice
T22945	11422-11423	.	denotes	.
T21857	11902-11908	VBN	denotes	Nested
T21859	11909-11912	NN	denotes	PCR
T21858	11913-11921	NN	denotes	strategy
T21860	11922-11925	IN	denotes	for
T21861	11926-11936	NN	denotes	genotyping
T21862	11937-11939	IN	denotes	of
T21863	11940-11956	JJ	denotes	pre-implantation
T21864	11957-11964	NNS	denotes	embryos
T21865	11964-11965	.	denotes	.
T21866	11965-12101	sentence	denotes	Due to the scarcity of extractable genetic material, a nested PCR strategy was developed in order to genotype pre-implantation embryos.
T21867	11966-11969	IN	denotes	Due
T21869	11970-11972	IN	denotes	to
T21870	11973-11976	DT	denotes	the
T21871	11977-11985	NN	denotes	scarcity
T21872	11986-11988	IN	denotes	of
T21873	11989-12000	JJ	denotes	extractable
T21875	12001-12008	JJ	denotes	genetic
T21874	12009-12017	NN	denotes	material
T21876	12017-12019	,	denotes	,
T21877	12019-12020	DT	denotes	a
T21879	12021-12027	VBN	denotes	nested
T21880	12028-12031	NN	denotes	PCR
T21878	12032-12040	NN	denotes	strategy
T21881	12041-12044	VBD	denotes	was
T21868	12045-12054	VBN	denotes	developed
T21882	12055-12057	IN	denotes	in
T21883	12058-12063	NN	denotes	order
T21884	12064-12066	TO	denotes	to
T21885	12067-12075	VB	denotes	genotype
T21886	12076-12092	JJ	denotes	pre-implantation
T21887	12093-12100	NNS	denotes	embryos
T21888	12100-12101	.	denotes	.
T21889	12101-12308	sentence	denotes	Separate reactions were used to amplify a 429 bp fragment from the wild-type allele and a 389 bp segment from the mutant allele, both spanning the 3' end of the short (upstream) arm of the targeting vector.
T21890	12102-12110	JJ	denotes	Separate
T21891	12111-12120	NNS	denotes	reactions
T21893	12121-12125	VBD	denotes	were
T21892	12126-12130	VBN	denotes	used
T21894	12131-12133	TO	denotes	to
T21895	12134-12141	VB	denotes	amplify
T21896	12142-12143	DT	denotes	a
T21898	12144-12147	CD	denotes	429
T21899	12148-12150	NN	denotes	bp
T21897	12151-12159	NN	denotes	fragment
T21900	12160-12164	IN	denotes	from
T21901	12165-12168	DT	denotes	the
T21903	12169-12173	JJ	denotes	wild
T21905	12173-12174	HYPH	denotes	-
T21904	12174-12178	NN	denotes	type
T21902	12179-12185	NN	denotes	allele
T21906	12186-12189	CC	denotes	and
T21907	12190-12191	DT	denotes	a
T21909	12192-12195	CD	denotes	389
T21910	12196-12198	NN	denotes	bp
T21908	12199-12206	NN	denotes	segment
T21911	12207-12211	IN	denotes	from
T21912	12212-12215	DT	denotes	the
T21914	12216-12222	NN	denotes	mutant
T21913	12223-12229	NN	denotes	allele
T21915	12229-12231	,	denotes	,
T21916	12231-12235	DT	denotes	both
T21917	12236-12244	VBG	denotes	spanning
T21918	12245-12248	DT	denotes	the
T21920	12249-12250	CD	denotes	3
T21921	12250-12251	SYM	denotes	'
T21919	12252-12255	NN	denotes	end
T21922	12256-12258	IN	denotes	of
T21923	12259-12262	DT	denotes	the
T21925	12263-12268	JJ	denotes	short
T21926	12269-12270	-LRB-	denotes	(
T21927	12270-12278	JJ	denotes	upstream
T21928	12278-12279	-RRB-	denotes	)
T21924	12280-12283	NN	denotes	arm
T21929	12284-12286	IN	denotes	of
T21930	12287-12290	DT	denotes	the
T21932	12291-12300	NN	denotes	targeting
T21931	12301-12307	NN	denotes	vector
T21933	12307-12308	.	denotes	.
T21934	12308-12446	sentence	denotes	In both reactions, a 213 bp sequence located within the short arm was co-amplified with the 'diagnostic' products as an internal control.
T21935	12309-12311	IN	denotes	In
T21937	12312-12316	DT	denotes	both
T21938	12317-12326	NNS	denotes	reactions
T21939	12326-12328	,	denotes	,
T21940	12328-12329	DT	denotes	a
T21942	12330-12333	CD	denotes	213
T21943	12334-12336	NN	denotes	bp
T21941	12337-12345	NN	denotes	sequence
T21944	12346-12353	VBN	denotes	located
T21945	12354-12360	IN	denotes	within
T21946	12361-12364	DT	denotes	the
T21948	12365-12370	JJ	denotes	short
T21947	12371-12374	NN	denotes	arm
T21949	12375-12378	VBD	denotes	was
T21936	12379-12391	VBN	denotes	co-amplified
T21950	12392-12396	IN	denotes	with
T21951	12397-12400	DT	denotes	the
T21953	12401-12402	``	denotes	'
T21954	12402-12412	JJ	denotes	diagnostic
T21955	12412-12413	''	denotes	'
T21952	12414-12422	NNS	denotes	products
T21956	12423-12425	IN	denotes	as
T21957	12426-12428	DT	denotes	an
T21959	12429-12437	JJ	denotes	internal
T21958	12438-12445	NN	denotes	control
T21960	12445-12446	.	denotes	.
T21961	12446-12584	sentence	denotes	The same sense primers were used to amplify 'diagnostic' sequences in both reactions, whereas the antisense primers were allele-specific.
T21962	12447-12450	DT	denotes	The
T21964	12451-12455	JJ	denotes	same
T21965	12456-12461	NN	denotes	sense
T21963	12462-12469	NNS	denotes	primers
T21967	12470-12474	VBD	denotes	were
T21966	12475-12479	VBN	denotes	used
T21968	12480-12482	TO	denotes	to
T21969	12483-12490	VB	denotes	amplify
T21970	12491-12492	``	denotes	'
T21971	12492-12502	JJ	denotes	diagnostic
T21973	12502-12503	''	denotes	'
T21972	12504-12513	NNS	denotes	sequences
T21974	12514-12516	IN	denotes	in
T21975	12517-12521	DT	denotes	both
T21976	12522-12531	NNS	denotes	reactions
T21977	12531-12533	,	denotes	,
T21978	12533-12540	IN	denotes	whereas
T21980	12541-12544	DT	denotes	the
T21982	12545-12554	NN	denotes	antisense
T21981	12555-12562	NNS	denotes	primers
T21979	12563-12567	VBD	denotes	were
T21983	12568-12574	NN	denotes	allele
T21985	12574-12575	HYPH	denotes	-
T21984	12575-12583	JJ	denotes	specific
T21986	12583-12584	.	denotes	.
T21987	12584-12690	sentence	denotes	The primers, represented by triangles, are depicted in two (nested) sets for each of the three reactions.
T21988	12585-12588	DT	denotes	The
T21989	12589-12596	NNS	denotes	primers
T21991	12596-12598	,	denotes	,
T21992	12598-12609	VBN	denotes	represented
T21993	12610-12612	IN	denotes	by
T21994	12613-12622	NNS	denotes	triangles
T21995	12622-12624	,	denotes	,
T21996	12624-12627	VBP	denotes	are
T21990	12628-12636	VBN	denotes	depicted
T21997	12637-12639	IN	denotes	in
T21998	12640-12643	CD	denotes	two
T22000	12644-12645	-LRB-	denotes	(
T22001	12645-12651	VBN	denotes	nested
T22002	12651-12652	-RRB-	denotes	)
T21999	12653-12657	NNS	denotes	sets
T22003	12658-12661	IN	denotes	for
T22004	12662-12666	DT	denotes	each
T22005	12667-12669	IN	denotes	of
T22006	12670-12673	DT	denotes	the
T22008	12674-12679	CD	denotes	three
T22007	12680-12689	NNS	denotes	reactions
T22009	12689-12690	.	denotes	.
T22010	12690-12907	sentence	denotes	Exons are represented by open, vertical rectangles, the PGK-Neo cassette by an open, horizontal rectangle, while thin vertical lines denote the boundaries of the short arm and the 5' end of the long (downstream) arm.
T22011	12691-12696	NNS	denotes	Exons
T22013	12697-12700	VBP	denotes	are
T22012	12701-12712	VBN	denotes	represented
T22015	12713-12715	IN	denotes	by
T22016	12716-12720	JJ	denotes	open
T22018	12720-12722	,	denotes	,
T22019	12722-12730	JJ	denotes	vertical
T22017	12731-12741	NNS	denotes	rectangles
T22020	12741-12743	,	denotes	,
T22021	12743-12746	DT	denotes	the
T22022	12747-12750	NN	denotes	PGK
T22024	12750-12751	HYPH	denotes	-
T22023	12751-12754	NN	denotes	Neo
T22014	12755-12763	NN	denotes	cassette
T22025	12764-12766	IN	denotes	by
T22026	12767-12769	DT	denotes	an
T22028	12770-12774	JJ	denotes	open
T22029	12774-12776	,	denotes	,
T22030	12776-12786	JJ	denotes	horizontal
T22027	12787-12796	NN	denotes	rectangle
T22031	12796-12798	,	denotes	,
T22032	12798-12803	IN	denotes	while
T22034	12804-12808	JJ	denotes	thin
T22036	12809-12817	JJ	denotes	vertical
T22035	12818-12823	NNS	denotes	lines
T22033	12824-12830	VBP	denotes	denote
T22037	12831-12834	DT	denotes	the
T22038	12835-12845	NNS	denotes	boundaries
T22039	12846-12848	IN	denotes	of
T22040	12849-12852	DT	denotes	the
T22042	12853-12858	JJ	denotes	short
T22041	12859-12862	NN	denotes	arm
T22043	12863-12866	CC	denotes	and
T22044	12867-12870	DT	denotes	the
T22046	12871-12872	CD	denotes	5
T22047	12872-12873	SYM	denotes	'
T22045	12874-12877	NN	denotes	end
T22048	12878-12880	IN	denotes	of
T22049	12881-12884	DT	denotes	the
T22051	12885-12889	JJ	denotes	long
T22052	12890-12891	-LRB-	denotes	(
T22053	12891-12901	JJ	denotes	downstream
T22054	12901-12902	-RRB-	denotes	)
T22050	12903-12906	NN	denotes	arm
T22055	12906-12907	.	denotes	.
T22056	12907-13046	sentence	denotes	A grey, horizontal rectangle delineates the segment of the wild-type allele that is replaced by the PGK-Neo cassette in the mutant allele.
T22057	12908-12909	DT	denotes	A
T22059	12910-12914	JJ	denotes	grey
T22060	12914-12916	,	denotes	,
T22061	12916-12926	JJ	denotes	horizontal
T22058	12927-12936	NN	denotes	rectangle
T22062	12937-12947	VBZ	denotes	delineates
T22063	12948-12951	DT	denotes	the
T22064	12952-12959	NN	denotes	segment
T22065	12960-12962	IN	denotes	of
T22066	12963-12966	DT	denotes	the
T22068	12967-12971	JJ	denotes	wild
T22070	12971-12972	HYPH	denotes	-
T22069	12972-12976	NN	denotes	type
T22067	12977-12983	NN	denotes	allele
T22071	12984-12988	WDT	denotes	that
T22073	12989-12991	VBZ	denotes	is
T22072	12992-13000	VBN	denotes	replaced
T22074	13001-13003	IN	denotes	by
T22075	13004-13007	DT	denotes	the
T22077	13008-13011	NN	denotes	PGK
T22079	13011-13012	HYPH	denotes	-
T22078	13012-13015	NN	denotes	Neo
T22076	13016-13024	NN	denotes	cassette
T22080	13025-13027	IN	denotes	in
T22081	13028-13031	DT	denotes	the
T22083	13032-13038	NN	denotes	mutant
T22082	13039-13045	NN	denotes	allele
T22084	13045-13046	.	denotes	.
T22640	13057-13067	NN	denotes	Genotyping
T22641	13068-13070	IN	denotes	of
T22642	13071-13087	JJ	denotes	pre-implantation
T22643	13088-13095	NNS	denotes	embryos
T22644	13095-13096	.	denotes	.
T22645	13096-13195	sentence	denotes	A nested PCR strategy was used to genotype embryos prior to implantation as described in Figure 4.
T22646	13097-13098	DT	denotes	A
T22648	13099-13105	VBN	denotes	nested
T22649	13106-13109	NN	denotes	PCR
T22647	13110-13118	NN	denotes	strategy
T22651	13119-13122	VBD	denotes	was
T22650	13123-13127	VBN	denotes	used
T22652	13128-13130	TO	denotes	to
T22653	13131-13139	VB	denotes	genotype
T22654	13140-13147	NNS	denotes	embryos
T22655	13148-13153	JJ	denotes	prior
T22656	13154-13156	IN	denotes	to
T22657	13157-13169	NN	denotes	implantation
T22658	13170-13172	IN	denotes	as
T22659	13173-13182	VBN	denotes	described
T22660	13183-13185	IN	denotes	in
T22661	13186-13192	NN	denotes	Figure
T22662	13193-13194	CD	denotes	4
T22663	13194-13195	.	denotes	.
T22664	13195-13298	sentence	denotes	Capn2+/- mice were mated and the date of fertilization established by the appearance of vaginal plugs.
T22665	13196-13201	NN	denotes	Capn2
T22667	13201-13202	SYM	denotes	+
T22668	13202-13203	HYPH	denotes	/
T22669	13203-13204	SYM	denotes	-
T22666	13205-13209	NNS	denotes	mice
T22671	13210-13214	VBD	denotes	were
T22670	13215-13220	VBN	denotes	mated
T22672	13221-13224	CC	denotes	and
T22673	13225-13228	DT	denotes	the
T22674	13229-13233	NN	denotes	date
T22676	13234-13236	IN	denotes	of
T22677	13237-13250	NN	denotes	fertilization
T22675	13251-13262	VBN	denotes	established
T22678	13263-13265	IN	denotes	by
T22679	13266-13269	DT	denotes	the
T22680	13270-13280	NN	denotes	appearance
T22681	13281-13283	IN	denotes	of
T22682	13284-13291	JJ	denotes	vaginal
T22683	13292-13297	NNS	denotes	plugs
T22684	13297-13298	.	denotes	.
T22685	13298-13409	sentence	denotes	Blastocyst (E3.5) or 8-cell embryos (E2.5) were flushed from the oviducts and then digested with proteinase K.
T22686	13299-13309	NN	denotes	Blastocyst
T22688	13310-13311	-LRB-	denotes	(
T22689	13311-13315	NN	denotes	E3.5
T22690	13315-13316	-RRB-	denotes	)
T22691	13317-13319	CC	denotes	or
T22692	13320-13321	CD	denotes	8
T22694	13321-13322	HYPH	denotes	-
T22693	13322-13326	NN	denotes	cell
T22687	13327-13334	NNS	denotes	embryos
T22696	13335-13336	-LRB-	denotes	(
T22697	13336-13340	NN	denotes	E2.5
T22698	13340-13341	-RRB-	denotes	)
T22699	13342-13346	VBD	denotes	were
T22695	13347-13354	VBN	denotes	flushed
T22700	13355-13359	IN	denotes	from
T22701	13360-13363	DT	denotes	the
T22702	13364-13372	NNS	denotes	oviducts
T22703	13373-13376	CC	denotes	and
T22704	13377-13381	RB	denotes	then
T22705	13382-13390	VBN	denotes	digested
T22706	13391-13395	IN	denotes	with
T22707	13396-13406	NN	denotes	proteinase
T22708	13407-13408	NN	denotes	K
T22709	13408-13409	.	denotes	.
T22710	13409-13646	sentence	denotes	In separate reactions segments found exclusively in either the wild-type or mutant alleles were co-amplified with an internal control sequence, located in the short (upstream) arm of the targeting vector, which is found in both alleles.
T22711	13410-13412	IN	denotes	In
T22713	13413-13421	JJ	denotes	separate
T22715	13422-13431	NNS	denotes	reactions
T22714	13432-13440	NNS	denotes	segments
T22716	13441-13446	VBN	denotes	found
T22717	13447-13458	RB	denotes	exclusively
T22718	13459-13461	IN	denotes	in
T22719	13462-13468	CC	denotes	either
T22721	13469-13472	DT	denotes	the
T22722	13473-13477	JJ	denotes	wild
T22723	13477-13478	HYPH	denotes	-
T22720	13478-13482	NN	denotes	type
T22724	13483-13485	CC	denotes	or
T22725	13486-13492	NN	denotes	mutant
T22726	13493-13500	NNS	denotes	alleles
T22727	13501-13505	VBD	denotes	were
T22712	13506-13518	VBN	denotes	co-amplified
T22728	13519-13523	IN	denotes	with
T22729	13524-13526	DT	denotes	an
T22731	13527-13535	JJ	denotes	internal
T22732	13536-13543	NN	denotes	control
T22730	13544-13552	NN	denotes	sequence
T22733	13552-13554	,	denotes	,
T22734	13554-13561	VBN	denotes	located
T22735	13562-13564	IN	denotes	in
T22736	13565-13568	DT	denotes	the
T22738	13569-13574	JJ	denotes	short
T22739	13575-13576	-LRB-	denotes	(
T22740	13576-13584	JJ	denotes	upstream
T22741	13584-13585	-RRB-	denotes	)
T22737	13586-13589	NN	denotes	arm
T22742	13590-13592	IN	denotes	of
T22743	13593-13596	DT	denotes	the
T22745	13597-13606	NN	denotes	targeting
T22744	13607-13613	NN	denotes	vector
T22746	13613-13615	,	denotes	,
T22747	13615-13620	WDT	denotes	which
T22749	13621-13623	VBZ	denotes	is
T22748	13624-13629	VBN	denotes	found
T22750	13630-13632	IN	denotes	in
T22751	13633-13637	DT	denotes	both
T22752	13638-13645	NNS	denotes	alleles
T22753	13645-13646	.	denotes	.
T22754	13646-13770	sentence	denotes	The final products were 429 bp for the wild-type allele, 389 bp for the mutant allele, and 213 bp for the internal control.
T22755	13647-13650	DT	denotes	The
T22757	13651-13656	JJ	denotes	final
T22756	13657-13665	NNS	denotes	products
T22758	13666-13670	VBD	denotes	were
T22759	13671-13674	CD	denotes	429
T22760	13675-13677	NN	denotes	bp
T22761	13678-13681	IN	denotes	for
T22762	13682-13685	DT	denotes	the
T22764	13686-13690	JJ	denotes	wild
T22766	13690-13691	HYPH	denotes	-
T22765	13691-13695	NN	denotes	type
T22763	13696-13702	NN	denotes	allele
T22767	13702-13704	,	denotes	,
T22768	13704-13707	CD	denotes	389
T22769	13708-13710	NN	denotes	bp
T22770	13711-13714	IN	denotes	for
T22771	13715-13718	DT	denotes	the
T22773	13719-13725	NN	denotes	mutant
T22772	13726-13732	NN	denotes	allele
T22774	13732-13734	,	denotes	,
T22775	13734-13737	CC	denotes	and
T22776	13738-13741	CD	denotes	213
T22777	13742-13744	NN	denotes	bp
T22778	13745-13748	IN	denotes	for
T22779	13749-13752	DT	denotes	the
T22781	13753-13761	JJ	denotes	internal
T22780	13762-13769	NN	denotes	control
T22782	13769-13770	.	denotes	.
T22783	13770-13846	sentence	denotes	(A) A representative example of the genotyping of blastocyst stage embryos.
T22784	13771-13772	-LRB-	denotes	(
T22785	13772-13773	LS	denotes	A
T22787	13773-13774	-RRB-	denotes	)
T22788	13775-13776	DT	denotes	A
T22789	13777-13791	JJ	denotes	representative
T22786	13792-13799	NN	denotes	example
T22790	13800-13802	IN	denotes	of
T22791	13803-13806	DT	denotes	the
T22792	13807-13817	NN	denotes	genotyping
T22793	13818-13820	IN	denotes	of
T22794	13821-13831	NN	denotes	blastocyst
T22796	13832-13837	NN	denotes	stage
T22795	13838-13845	NNS	denotes	embryos
T22797	13845-13846	.	denotes	.
T22798	13846-13980	sentence	denotes	Embryos #1, 2, 4, 5, and 6 were Capn2+/- whereas embryos #3 and #7 were Capn2+/+, denoted by the absence of the 389 bp mutant signal.
T22799	13847-13854	NNS	denotes	Embryos
T22801	13855-13856	SYM	denotes	#
T22800	13856-13857	CD	denotes	1
T22803	13857-13859	,	denotes	,
T22804	13859-13860	CD	denotes	2
T22805	13860-13862	,	denotes	,
T22806	13862-13863	CD	denotes	4
T22807	13863-13865	,	denotes	,
T22808	13865-13866	CD	denotes	5
T22809	13866-13868	,	denotes	,
T22810	13868-13871	CC	denotes	and
T22811	13872-13873	CD	denotes	6
T22802	13874-13878	VBD	denotes	were
T22812	13879-13884	NN	denotes	Capn2
T22813	13884-13885	SYM	denotes	+
T22814	13885-13886	HYPH	denotes	/
T22815	13886-13887	SYM	denotes	-
T22816	13888-13895	IN	denotes	whereas
T22818	13896-13903	NNS	denotes	embryos
T22820	13904-13905	SYM	denotes	#
T22819	13905-13906	CD	denotes	3
T22821	13907-13910	CC	denotes	and
T22822	13911-13912	SYM	denotes	#
T22823	13912-13913	CD	denotes	7
T22817	13914-13918	VBD	denotes	were
T22824	13919-13924	NN	denotes	Capn2
T22825	13924-13925	SYM	denotes	+
T22826	13925-13926	HYPH	denotes	/
T22827	13926-13927	SYM	denotes	+
T22828	13927-13929	,	denotes	,
T22829	13929-13936	VBN	denotes	denoted
T22830	13937-13939	IN	denotes	by
T22831	13940-13943	DT	denotes	the
T22832	13944-13951	NN	denotes	absence
T22833	13952-13954	IN	denotes	of
T22834	13955-13958	DT	denotes	the
T22836	13959-13962	CD	denotes	389
T22837	13963-13965	NN	denotes	bp
T22838	13966-13972	NN	denotes	mutant
T22835	13973-13979	NN	denotes	signal
T22839	13979-13980	.	denotes	.
T22840	13980-14041	sentence	denotes	(B) An example of the genotyping of 8-cell embryos is shown.
T22841	13981-13982	-LRB-	denotes	(
T22842	13982-13983	LS	denotes	B
T22844	13983-13984	-RRB-	denotes	)
T22845	13985-13987	DT	denotes	An
T22846	13988-13995	NN	denotes	example
T22847	13996-13998	IN	denotes	of
T22848	13999-14002	DT	denotes	the
T22849	14003-14013	NN	denotes	genotyping
T22850	14014-14016	IN	denotes	of
T22851	14017-14018	CD	denotes	8
T22853	14018-14019	HYPH	denotes	-
T22852	14019-14023	NN	denotes	cell
T22854	14024-14031	NNS	denotes	embryos
T22855	14032-14034	VBZ	denotes	is
T22843	14035-14040	VBN	denotes	shown
T22856	14040-14041	.	denotes	.
T22857	14041-14166	sentence	denotes	Embryos #1, 2, 3, 5, and 6 were Capn2+/- while embryo #4 was Capn2-/-, marked by the absence of the 429 bp wild-type signal.
T22858	14042-14049	NNS	denotes	Embryos
T22860	14050-14051	SYM	denotes	#
T22859	14051-14052	CD	denotes	1
T22862	14052-14054	,	denotes	,
T22863	14054-14055	CD	denotes	2
T22864	14055-14057	,	denotes	,
T22865	14057-14058	CD	denotes	3
T22866	14058-14060	,	denotes	,
T22867	14060-14061	CD	denotes	5
T22868	14061-14063	,	denotes	,
T22869	14063-14066	CC	denotes	and
T22870	14067-14068	CD	denotes	6
T22861	14069-14073	VBD	denotes	were
T22871	14074-14079	NN	denotes	Capn2
T22872	14079-14080	SYM	denotes	+
T22873	14080-14081	HYPH	denotes	/
T22874	14081-14082	SYM	denotes	-
T22875	14083-14088	IN	denotes	while
T22877	14089-14095	NN	denotes	embryo
T22878	14096-14097	SYM	denotes	#
T22879	14097-14098	CD	denotes	4
T22876	14099-14102	VBD	denotes	was
T22880	14103-14108	NN	denotes	Capn2
T22881	14108-14109	SYM	denotes	-
T22882	14109-14110	HYPH	denotes	/
T22883	14110-14111	SYM	denotes	-
T22884	14111-14113	,	denotes	,
T22885	14113-14119	VBN	denotes	marked
T22886	14120-14122	IN	denotes	by
T22887	14123-14126	DT	denotes	the
T22888	14127-14134	NN	denotes	absence
T22889	14135-14137	IN	denotes	of
T22890	14138-14141	DT	denotes	the
T22892	14142-14145	CD	denotes	429
T22893	14146-14148	NN	denotes	bp
T22894	14149-14153	JJ	denotes	wild
T22896	14153-14154	HYPH	denotes	-
T22895	14154-14158	NN	denotes	type
T22891	14159-14165	NN	denotes	signal
T22897	14165-14166	.	denotes	.
T22898	14166-14207	sentence	denotes	(M) denotes the molecular weight marker.
T22899	14167-14168	-LRB-	denotes	(
T22900	14168-14169	NN	denotes	M
T22902	14169-14170	-RRB-	denotes	)
T22901	14171-14178	VBZ	denotes	denotes
T22903	14179-14182	DT	denotes	the
T22905	14183-14192	JJ	denotes	molecular
T22906	14193-14199	NN	denotes	weight
T22904	14200-14206	NN	denotes	marker
T22907	14206-14207	.	denotes	.
T7006	14212-14222	NN	denotes	genotyping
T7008	14223-14230	VBZ	denotes	results
T7009	14231-14234	IN	denotes	for
T7010	14235-14244	NNS	denotes	weanlings
T7011	14244-14246	,	denotes	,
T7012	14246-14263	JJ	denotes	post-implantation
T7014	14263-14265	,	denotes	,
T7015	14265-14268	CC	denotes	and
T7016	14269-14285	JJ	denotes	pre-implantation
T7013	14286-14293	NNS	denotes	embryos
T7017	14294-14297	VBP	denotes	are
T7007	14298-14303	VBN	denotes	shown
T7018	14304-14306	IN	denotes	in
T7019	14307-14312	NN	denotes	Table
T7020	14313-14314	CD	denotes	2
T7021	14314-14315	.	denotes	.
T7022	14315-14468	sentence	denotes	Curiously, the ratio of Capn2+/+ to Capn2+/-animals from inbred heterozygous intercrosses was substantially less than the predicted 1:2 Mendelian ratio.
T7023	14316-14325	RB	denotes	Curiously
T7025	14325-14327	,	denotes	,
T7026	14327-14330	DT	denotes	the
T7027	14331-14336	NN	denotes	ratio
T7028	14337-14339	IN	denotes	of
T7029	14340-14345	NN	denotes	Capn2
T7030	14345-14346	SYM	denotes	+
T7031	14346-14347	HYPH	denotes	/
T7032	14347-14348	SYM	denotes	+
T7033	14349-14351	IN	denotes	to
T7034	14352-14357	NN	denotes	Capn2
T7036	14357-14358	SYM	denotes	+
T7037	14358-14359	HYPH	denotes	/
T7038	14359-14360	SYM	denotes	-
T7035	14360-14367	NNS	denotes	animals
T7039	14368-14372	IN	denotes	from
T7040	14373-14379	JJ	denotes	inbred
T7042	14380-14392	JJ	denotes	heterozygous
T7041	14393-14405	NNS	denotes	intercrosses
T7024	14406-14409	VBD	denotes	was
T7043	14410-14423	RB	denotes	substantially
T7044	14424-14428	JJR	denotes	less
T7045	14429-14433	IN	denotes	than
T7046	14434-14437	DT	denotes	the
T7048	14438-14447	VBN	denotes	predicted
T7049	14448-14449	CD	denotes	1
T7050	14449-14450	SYM	denotes	:
T7051	14450-14451	CD	denotes	2
T7052	14452-14461	JJ	denotes	Mendelian
T7047	14462-14467	NN	denotes	ratio
T7053	14467-14468	.	denotes	.
T7054	14468-14611	sentence	denotes	In a group of 199 animals derived from heterozygote breeding (33 litters), 23 Capn2+/+ (11.6%) and 176 (88.4%) Capn2+/- animals were observed.
T7055	14469-14471	IN	denotes	In
T7057	14472-14473	DT	denotes	a
T7058	14474-14479	NN	denotes	group
T7059	14480-14482	IN	denotes	of
T7060	14483-14486	CD	denotes	199
T7061	14487-14494	NNS	denotes	animals
T7062	14495-14502	VBN	denotes	derived
T7063	14503-14507	IN	denotes	from
T7064	14508-14520	NN	denotes	heterozygote
T7065	14521-14529	NN	denotes	breeding
T7066	14530-14531	-LRB-	denotes	(
T7068	14531-14533	CD	denotes	33
T7067	14534-14541	NNS	denotes	litters
T7069	14541-14542	-RRB-	denotes	)
T7070	14542-14544	,	denotes	,
T7071	14544-14546	CD	denotes	23
T7072	14547-14552	NN	denotes	Capn2
T7073	14552-14553	SYM	denotes	+
T7074	14553-14554	HYPH	denotes	/
T7075	14554-14555	SYM	denotes	+
T7076	14556-14557	-LRB-	denotes	(
T7078	14557-14561	CD	denotes	11.6
T7077	14561-14562	NN	denotes	%
T7079	14562-14563	-RRB-	denotes	)
T7080	14564-14567	CC	denotes	and
T7081	14568-14571	CD	denotes	176
T7083	14572-14573	-LRB-	denotes	(
T7085	14573-14577	CD	denotes	88.4
T7084	14577-14578	NN	denotes	%
T7086	14578-14579	-RRB-	denotes	)
T7082	14580-14585	NN	denotes	Capn2
T7088	14585-14586	SYM	denotes	+
T7089	14586-14587	HYPH	denotes	/
T7090	14587-14588	SYM	denotes	-
T7087	14589-14596	NNS	denotes	animals
T7091	14597-14601	VBD	denotes	were
T7056	14602-14610	VBN	denotes	observed
T7092	14610-14611	.	denotes	.
T7093	14611-14860	sentence	denotes	The ratio of Capn2+/+ to Capn2+/- animals among males (14% to 82%) or females (13% to 90%) was essentially the same is it was for the combined population, and there were an average of six pups per litter, which is normal for this background strain.
T7094	14612-14615	DT	denotes	The
T7095	14616-14621	NN	denotes	ratio
T7097	14622-14624	IN	denotes	of
T7098	14625-14630	NN	denotes	Capn2
T7099	14630-14631	SYM	denotes	+
T7100	14631-14632	HYPH	denotes	/
T7101	14632-14633	SYM	denotes	+
T7102	14634-14636	IN	denotes	to
T7103	14637-14642	NN	denotes	Capn2
T7105	14642-14643	SYM	denotes	+
T7106	14643-14644	HYPH	denotes	/
T7107	14644-14645	SYM	denotes	-
T7104	14646-14653	NNS	denotes	animals
T7108	14654-14659	IN	denotes	among
T7109	14660-14665	NNS	denotes	males
T7110	14666-14667	-LRB-	denotes	(
T7112	14667-14669	CD	denotes	14
T7111	14669-14670	NN	denotes	%
T7113	14671-14673	IN	denotes	to
T7114	14674-14676	CD	denotes	82
T7115	14676-14677	NN	denotes	%
T7116	14677-14678	-RRB-	denotes	)
T7117	14679-14681	CC	denotes	or
T7118	14682-14689	NNS	denotes	females
T7119	14690-14691	-LRB-	denotes	(
T7121	14691-14693	CD	denotes	13
T7120	14693-14694	NN	denotes	%
T7122	14695-14697	IN	denotes	to
T7123	14698-14700	CD	denotes	90
T7124	14700-14701	NN	denotes	%
T7125	14701-14702	-RRB-	denotes	)
T7096	14703-14706	VBD	denotes	was
T7126	14707-14718	RB	denotes	essentially
T7127	14719-14722	DT	denotes	the
T7128	14723-14727	JJ	denotes	same
T7129	14728-14730	IN	denotes	is
T7131	14731-14733	PRP	denotes	it
T7130	14734-14737	VBD	denotes	was
T7132	14738-14741	IN	denotes	for
T7133	14742-14745	DT	denotes	the
T7135	14746-14754	VBN	denotes	combined
T7134	14755-14765	NN	denotes	population
T7136	14765-14767	,	denotes	,
T7137	14767-14770	CC	denotes	and
T7138	14771-14776	EX	denotes	there
T7139	14777-14781	VBD	denotes	were
T7140	14782-14784	DT	denotes	an
T7141	14785-14792	NN	denotes	average
T7142	14793-14795	IN	denotes	of
T7143	14796-14799	CD	denotes	six
T7144	14800-14804	NNS	denotes	pups
T7145	14805-14808	IN	denotes	per
T7146	14809-14815	NN	denotes	litter
T7147	14815-14817	,	denotes	,
T7148	14817-14822	WDT	denotes	which
T7149	14823-14825	VBZ	denotes	is
T7150	14826-14832	JJ	denotes	normal
T7151	14833-14836	IN	denotes	for
T7152	14837-14841	DT	denotes	this
T7154	14842-14852	NN	denotes	background
T7153	14853-14859	NN	denotes	strain
T7155	14859-14860	.	denotes	.
T7156	14860-14994	sentence	denotes	A larger than expected proportion Capn2+/- animals was also evident in both post-implantation and pre-implantation embryos (Table 2).
T7157	14861-14862	DT	denotes	A
T7159	14863-14869	JJR	denotes	larger
T7160	14870-14874	IN	denotes	than
T7161	14875-14883	VBN	denotes	expected
T7158	14884-14894	NN	denotes	proportion
T7163	14895-14900	NN	denotes	Capn2
T7165	14900-14901	SYM	denotes	+
T7166	14901-14902	HYPH	denotes	/
T7167	14902-14903	SYM	denotes	-
T7164	14904-14911	NNS	denotes	animals
T7162	14912-14915	VBD	denotes	was
T7168	14916-14920	RB	denotes	also
T7169	14921-14928	JJ	denotes	evident
T7170	14929-14931	IN	denotes	in
T7171	14932-14936	CC	denotes	both
T7173	14937-14954	JJ	denotes	post-implantation
T7174	14955-14958	CC	denotes	and
T7175	14959-14975	JJ	denotes	pre-implantation
T7172	14976-14983	NNS	denotes	embryos
T7176	14984-14985	-LRB-	denotes	(
T7177	14985-14990	NN	denotes	Table
T7178	14991-14992	CD	denotes	2
T7179	14992-14993	-RRB-	denotes	)
T7180	14993-14994	.	denotes	.
T7181	14994-15172	sentence	denotes	Interestingly, a similar over-representation of heterozygous animals was also reported in one of the Capn4 transgenic lines, though the genotype skewing was not as extreme [32].
T7182	14995-15008	RB	denotes	Interestingly
T7184	15008-15010	,	denotes	,
T7185	15010-15011	DT	denotes	a
T7187	15012-15019	JJ	denotes	similar
T7186	15020-15039	NN	denotes	over-representation
T7188	15040-15042	IN	denotes	of
T7189	15043-15055	JJ	denotes	heterozygous
T7190	15056-15063	NNS	denotes	animals
T7191	15064-15067	VBD	denotes	was
T7192	15068-15072	RB	denotes	also
T7183	15073-15081	VBN	denotes	reported
T7193	15082-15084	IN	denotes	in
T7194	15085-15088	CD	denotes	one
T7195	15089-15091	IN	denotes	of
T7196	15092-15095	DT	denotes	the
T7198	15096-15101	NN	denotes	Capn4
T7199	15102-15112	JJ	denotes	transgenic
T7197	15113-15118	NNS	denotes	lines
T7200	15118-15120	,	denotes	,
T7201	15120-15126	IN	denotes	though
T7203	15127-15130	DT	denotes	the
T7205	15131-15139	NN	denotes	genotype
T7204	15140-15147	NN	denotes	skewing
T7202	15148-15151	VBD	denotes	was
T7206	15152-15155	RB	denotes	not
T7207	15156-15158	IN	denotes	as
T7208	15159-15166	JJ	denotes	extreme
T7209	15167-15168	-LRB-	denotes	[
T7210	15168-15170	CD	denotes	32
T7211	15170-15171	-RRB-	denotes	]
T7212	15171-15172	.	denotes	.
T7213	15172-15301	sentence	denotes	Crosses between Capn2+/+ and Capn2+/- animals also resulted in a greater than expected proportion of Capn2+/- animals (Table 2).
T7214	15173-15180	NNS	denotes	Crosses
T7216	15181-15188	IN	denotes	between
T7217	15189-15194	NN	denotes	Capn2
T7219	15194-15195	SYM	denotes	+
T7220	15195-15196	HYPH	denotes	/
T7221	15196-15197	SYM	denotes	+
T7222	15198-15201	CC	denotes	and
T7223	15202-15207	NN	denotes	Capn2
T7224	15207-15208	SYM	denotes	+
T7225	15208-15209	HYPH	denotes	/
T7226	15209-15210	SYM	denotes	-
T7218	15211-15218	NNS	denotes	animals
T7227	15219-15223	RB	denotes	also
T7215	15224-15232	VBD	denotes	resulted
T7228	15233-15235	IN	denotes	in
T7229	15236-15237	DT	denotes	a
T7231	15238-15245	JJR	denotes	greater
T7232	15246-15250	IN	denotes	than
T7233	15251-15259	VBN	denotes	expected
T7230	15260-15270	NN	denotes	proportion
T7234	15271-15273	IN	denotes	of
T7235	15274-15279	NN	denotes	Capn2
T7237	15279-15280	SYM	denotes	+
T7238	15280-15281	HYPH	denotes	/
T7239	15281-15282	SYM	denotes	-
T7236	15283-15290	NNS	denotes	animals
T7240	15291-15292	-LRB-	denotes	(
T7241	15292-15297	NN	denotes	Table
T7242	15298-15299	CD	denotes	2
T7243	15299-15300	-RRB-	denotes	)
T7244	15300-15301	.	denotes	.
T7245	15301-15457	sentence	denotes	An even higher degree of Capn2+/- over-representation was seen when the mutant allele came the mother (73%) compared to when it came from the father (59%).
T7246	15302-15304	DT	denotes	An
T7248	15305-15309	RB	denotes	even
T7249	15310-15316	JJR	denotes	higher
T7247	15317-15323	NN	denotes	degree
T7251	15324-15326	IN	denotes	of
T7252	15327-15332	NN	denotes	Capn2
T7254	15332-15333	SYM	denotes	+
T7255	15333-15334	HYPH	denotes	/
T7256	15334-15335	SYM	denotes	-
T7253	15336-15355	NN	denotes	over-representation
T7257	15356-15359	VBD	denotes	was
T7250	15360-15364	VBN	denotes	seen
T7258	15365-15369	WRB	denotes	when
T7260	15370-15373	DT	denotes	the
T7262	15374-15380	NN	denotes	mutant
T7261	15381-15387	NN	denotes	allele
T7259	15388-15392	VBD	denotes	came
T7263	15393-15396	DT	denotes	the
T7264	15397-15403	NN	denotes	mother
T7265	15404-15405	-LRB-	denotes	(
T7267	15405-15407	CD	denotes	73
T7266	15407-15408	NN	denotes	%
T7268	15408-15409	-RRB-	denotes	)
T7269	15410-15418	VBN	denotes	compared
T7270	15419-15421	IN	denotes	to
T7271	15422-15426	WRB	denotes	when
T7273	15427-15429	PRP	denotes	it
T7272	15430-15434	VBD	denotes	came
T7274	15435-15439	IN	denotes	from
T7275	15440-15443	DT	denotes	the
T7276	15444-15450	NN	denotes	father
T7277	15451-15452	-LRB-	denotes	(
T7279	15452-15454	CD	denotes	59
T7278	15454-15455	NN	denotes	%
T7280	15455-15456	-RRB-	denotes	)
T7281	15456-15457	.	denotes	.
T7282	15457-15784	sentence	denotes	In these crosses the ratios of Capn2+/+ to Capn2+/- animals among males or females compared well with the ratio in the combined populations; 77% of males and 69% of females were Capn2+/- when the mutant allele came from the mother, and 62% of males and 55% of females were Capn2+/- when the mutant allele came from the father.
T7283	15458-15460	IN	denotes	In
T7285	15461-15466	DT	denotes	these
T7286	15467-15474	NNS	denotes	crosses
T7287	15475-15478	DT	denotes	the
T7288	15479-15485	NNS	denotes	ratios
T7289	15486-15488	IN	denotes	of
T7290	15489-15494	NN	denotes	Capn2
T7291	15494-15495	SYM	denotes	+
T7292	15495-15496	HYPH	denotes	/
T7293	15496-15497	SYM	denotes	+
T7294	15498-15500	IN	denotes	to
T7295	15501-15506	NN	denotes	Capn2
T7297	15506-15507	SYM	denotes	+
T7298	15507-15508	HYPH	denotes	/
T7299	15508-15509	SYM	denotes	-
T7296	15510-15517	NNS	denotes	animals
T7300	15518-15523	IN	denotes	among
T7301	15524-15529	NNS	denotes	males
T7302	15530-15532	CC	denotes	or
T7303	15533-15540	NNS	denotes	females
T7284	15541-15549	VBN	denotes	compared
T7304	15550-15554	RB	denotes	well
T7305	15555-15559	IN	denotes	with
T7306	15560-15563	DT	denotes	the
T7307	15564-15569	NN	denotes	ratio
T7308	15570-15572	IN	denotes	in
T7309	15573-15576	DT	denotes	the
T7311	15577-15585	VBN	denotes	combined
T7310	15586-15597	NNS	denotes	populations
T7312	15597-15598	:	denotes	;
T7313	15599-15601	CD	denotes	77
T7314	15601-15602	NN	denotes	%
T7316	15603-15605	IN	denotes	of
T7317	15606-15611	NNS	denotes	males
T7318	15612-15615	CC	denotes	and
T7319	15616-15618	CD	denotes	69
T7320	15618-15619	NN	denotes	%
T7321	15620-15622	IN	denotes	of
T7322	15623-15630	NNS	denotes	females
T7315	15631-15635	VBD	denotes	were
T7323	15636-15641	NN	denotes	Capn2
T7324	15641-15642	SYM	denotes	+
T7325	15642-15643	HYPH	denotes	/
T7326	15643-15644	SYM	denotes	-
T7327	15645-15649	WRB	denotes	when
T7329	15650-15653	DT	denotes	the
T7331	15654-15660	NN	denotes	mutant
T7330	15661-15667	NN	denotes	allele
T7328	15668-15672	VBD	denotes	came
T7332	15673-15677	IN	denotes	from
T7333	15678-15681	DT	denotes	the
T7334	15682-15688	NN	denotes	mother
T7335	15688-15690	,	denotes	,
T7336	15690-15693	CC	denotes	and
T7337	15694-15696	CD	denotes	62
T7338	15696-15697	NN	denotes	%
T7340	15698-15700	IN	denotes	of
T7341	15701-15706	NNS	denotes	males
T7342	15707-15710	CC	denotes	and
T7343	15711-15713	CD	denotes	55
T7344	15713-15714	NN	denotes	%
T7345	15715-15717	IN	denotes	of
T7346	15718-15725	NNS	denotes	females
T7339	15726-15730	VBD	denotes	were
T7347	15731-15736	NN	denotes	Capn2
T7348	15736-15737	SYM	denotes	+
T7349	15737-15738	HYPH	denotes	/
T7350	15738-15739	SYM	denotes	-
T7351	15740-15744	WRB	denotes	when
T7353	15745-15748	DT	denotes	the
T7355	15749-15755	NN	denotes	mutant
T7354	15756-15762	NN	denotes	allele
T7352	15763-15767	VBD	denotes	came
T7356	15768-15772	IN	denotes	from
T7357	15773-15776	DT	denotes	the
T7358	15777-15783	NN	denotes	father
T7359	15783-15784	.	denotes	.
T7704	15786-15795	VBN	denotes	Attempted
T7705	15796-15806	NN	denotes	generation
T7706	15807-15809	IN	denotes	of
T7707	15810-15815	NN	denotes	Capn2
T7709	15815-15816	SYM	denotes	-
T7710	15816-15817	HYPH	denotes	/
T7711	15817-15818	SYM	denotes	-
T7712	15819-15821	NN	denotes	ES
T7708	15822-15827	NNS	denotes	cells
T7713	15827-15980	sentence	denotes	Capn2+/- ES cells were subjected to clonal selection in the presence of 2 mg/mL G418 in attempts to generate homozygous mutant cells by gene conversion.
T7714	15828-15833	NN	denotes	Capn2
T7716	15833-15834	SYM	denotes	+
T7717	15834-15835	HYPH	denotes	/
T7718	15835-15836	SYM	denotes	-
T7719	15837-15839	NN	denotes	ES
T7715	15840-15845	NNS	denotes	cells
T7721	15846-15850	VBD	denotes	were
T7720	15851-15860	VBN	denotes	subjected
T7722	15861-15863	IN	denotes	to
T7723	15864-15870	JJ	denotes	clonal
T7724	15871-15880	NN	denotes	selection
T7725	15881-15883	IN	denotes	in
T7726	15884-15887	DT	denotes	the
T7727	15888-15896	NN	denotes	presence
T7728	15897-15899	IN	denotes	of
T7729	15900-15901	CD	denotes	2
T7730	15902-15904	NN	denotes	mg
T7732	15904-15905	SYM	denotes	/
T7733	15905-15907	NN	denotes	mL
T7731	15908-15912	NN	denotes	G418
T7734	15913-15915	IN	denotes	in
T7735	15916-15924	NNS	denotes	attempts
T7736	15925-15927	TO	denotes	to
T7737	15928-15936	VB	denotes	generate
T7738	15937-15947	JJ	denotes	homozygous
T7740	15948-15954	NN	denotes	mutant
T7739	15955-15960	NNS	denotes	cells
T7741	15961-15963	IN	denotes	by
T7742	15964-15968	NN	denotes	gene
T7743	15969-15979	NN	denotes	conversion
T7744	15979-15980	.	denotes	.
T7745	15980-16109	sentence	denotes	This procedure has been extensively applied to targeted ES cells and was readily achieved in the case of Capn4+/- ES cells [31].
T7746	15981-15985	DT	denotes	This
T7747	15986-15995	NN	denotes	procedure
T7749	15996-15999	VBZ	denotes	has
T7750	16000-16004	VBN	denotes	been
T7751	16005-16016	RB	denotes	extensively
T7748	16017-16024	VBN	denotes	applied
T7752	16025-16027	IN	denotes	to
T7753	16028-16036	VBN	denotes	targeted
T7755	16037-16039	NN	denotes	ES
T7754	16040-16045	NNS	denotes	cells
T7756	16046-16049	CC	denotes	and
T7757	16050-16053	VBD	denotes	was
T7759	16054-16061	RB	denotes	readily
T7758	16062-16070	VBN	denotes	achieved
T7760	16071-16073	IN	denotes	in
T7761	16074-16077	DT	denotes	the
T7762	16078-16082	NN	denotes	case
T7763	16083-16085	IN	denotes	of
T7764	16086-16091	NN	denotes	Capn4
T7766	16091-16092	SYM	denotes	+
T7767	16092-16093	HYPH	denotes	/
T7768	16093-16094	SYM	denotes	-
T7769	16095-16097	NN	denotes	ES
T7765	16098-16103	NNS	denotes	cells
T7770	16104-16105	-LRB-	denotes	[
T7771	16105-16107	CD	denotes	31
T7772	16107-16108	-RRB-	denotes	]
T7773	16108-16109	.	denotes	.
T7774	16109-16208	sentence	denotes	In this case, however, no Capn2-/- ES cells were isolated in screens of 126 drug-resistant clones.
T7775	16110-16112	IN	denotes	In
T7777	16113-16117	DT	denotes	this
T7778	16118-16122	NN	denotes	case
T7779	16122-16124	,	denotes	,
T7780	16124-16131	RB	denotes	however
T7781	16131-16133	,	denotes	,
T7782	16133-16135	DT	denotes	no
T7784	16136-16141	NN	denotes	Capn2
T7785	16141-16142	SYM	denotes	-
T7786	16142-16143	HYPH	denotes	/
T7787	16143-16144	SYM	denotes	-
T7788	16145-16147	NN	denotes	ES
T7783	16148-16153	NNS	denotes	cells
T7789	16154-16158	VBD	denotes	were
T7776	16159-16167	VBN	denotes	isolated
T7790	16168-16170	IN	denotes	in
T7791	16171-16178	NNS	denotes	screens
T7792	16179-16181	IN	denotes	of
T7793	16182-16185	CD	denotes	126
T7795	16186-16190	NN	denotes	drug
T7797	16190-16191	HYPH	denotes	-
T7796	16191-16200	JJ	denotes	resistant
T7794	16201-16207	NNS	denotes	clones
T7798	16207-16208	.	denotes	.
T7799	16208-16444	sentence	denotes	The inability to isolate Capn2-/- ES cells, in concert with the absence of Capn2-/- embryos beyond the 8-cell stage, suggested that m-calpain activity might be essential for cell viability or the establishment of viable ES cell clones.
T7800	16209-16212	DT	denotes	The
T7801	16213-16222	NN	denotes	inability
T7803	16223-16225	TO	denotes	to
T7804	16226-16233	VB	denotes	isolate
T7805	16234-16239	NN	denotes	Capn2
T7807	16239-16240	SYM	denotes	-
T7808	16240-16241	HYPH	denotes	/
T7809	16241-16242	SYM	denotes	-
T7810	16243-16245	NN	denotes	ES
T7806	16246-16251	NNS	denotes	cells
T7811	16251-16253	,	denotes	,
T7812	16253-16255	IN	denotes	in
T7813	16256-16263	NN	denotes	concert
T7814	16264-16268	IN	denotes	with
T7815	16269-16272	DT	denotes	the
T7816	16273-16280	NN	denotes	absence
T7817	16281-16283	IN	denotes	of
T7818	16284-16289	NN	denotes	Capn2
T7820	16289-16290	SYM	denotes	-
T7821	16290-16291	HYPH	denotes	/
T7822	16291-16292	SYM	denotes	-
T7819	16293-16300	NNS	denotes	embryos
T7823	16301-16307	IN	denotes	beyond
T7824	16308-16311	DT	denotes	the
T7826	16312-16313	CD	denotes	8
T7828	16313-16314	HYPH	denotes	-
T7827	16314-16318	NN	denotes	cell
T7825	16319-16324	NN	denotes	stage
T7829	16324-16326	,	denotes	,
T7802	16326-16335	VBD	denotes	suggested
T7830	16336-16340	IN	denotes	that
T7832	16341-16342	NN	denotes	m
T7834	16342-16343	HYPH	denotes	-
T7833	16343-16350	NN	denotes	calpain
T7835	16351-16359	NN	denotes	activity
T7836	16360-16365	MD	denotes	might
T7831	16366-16368	VB	denotes	be
T7837	16369-16378	JJ	denotes	essential
T7838	16379-16382	IN	denotes	for
T7839	16383-16387	NN	denotes	cell
T7840	16388-16397	NN	denotes	viability
T7841	16398-16400	CC	denotes	or
T7842	16401-16404	DT	denotes	the
T7843	16405-16418	NN	denotes	establishment
T7844	16419-16421	IN	denotes	of
T7845	16422-16428	JJ	denotes	viable
T7847	16429-16431	NN	denotes	ES
T7848	16432-16436	NN	denotes	cell
T7846	16437-16443	NNS	denotes	clones
T7849	16443-16444	.	denotes	.
T12816	16457-16465	IN	denotes	Although
T12818	16466-16473	NN	denotes	calpain
T12819	16474-16482	NN	denotes	activity
T12820	16483-16486	VBD	denotes	was
T12821	16487-16492	RB	denotes	first
T12817	16493-16503	VBN	denotes	identified
T12823	16504-16508	CD	denotes	four
T12824	16509-16516	NNS	denotes	decades
T12825	16517-16520	RB	denotes	ago
T12826	16520-16522	,	denotes	,
T12827	16522-16523	DT	denotes	a
T12829	16524-16529	JJ	denotes	clear
T12828	16530-16543	NN	denotes	understanding
T12830	16544-16546	IN	denotes	of
T12831	16547-16550	DT	denotes	the
T12832	16551-16561	NNS	denotes	substrates
T12833	16562-16565	CC	denotes	and
T12834	16566-16575	NNS	denotes	functions
T12835	16576-16578	IN	denotes	of
T12836	16579-16582	DT	denotes	the
T12837	16583-16590	NNS	denotes	enzymes
T12838	16591-16594	VBZ	denotes	has
T12822	16595-16603	VBN	denotes	remained
T12839	16604-16611	JJ	denotes	elusive
T12840	16611-16612	.	denotes	.
T12841	16612-16759	sentence	denotes	In large part, this has been due to the lack of inhibitors capable of specifically down-regulating the calpains without affecting other proteases.
T12842	16613-16615	IN	denotes	In
T12844	16616-16621	JJ	denotes	large
T12845	16622-16626	NN	denotes	part
T12846	16626-16628	,	denotes	,
T12847	16628-16632	DT	denotes	this
T12848	16633-16636	VBZ	denotes	has
T12843	16637-16641	VBN	denotes	been
T12849	16642-16645	IN	denotes	due
T12850	16646-16648	IN	denotes	to
T12851	16649-16652	DT	denotes	the
T12852	16653-16657	NN	denotes	lack
T12853	16658-16660	IN	denotes	of
T12854	16661-16671	NNS	denotes	inhibitors
T12855	16672-16679	JJ	denotes	capable
T12856	16680-16682	IN	denotes	of
T12857	16683-16695	RB	denotes	specifically
T12859	16696-16700	RB	denotes	down
T12860	16700-16701	HYPH	denotes	-
T12858	16701-16711	VBG	denotes	regulating
T12861	16712-16715	DT	denotes	the
T12862	16716-16724	NNS	denotes	calpains
T12863	16725-16732	IN	denotes	without
T12864	16733-16742	VBG	denotes	affecting
T12865	16743-16748	JJ	denotes	other
T12866	16749-16758	NNS	denotes	proteases
T12867	16758-16759	.	denotes	.
T12868	16759-16955	sentence	denotes	In the past decade, the story has been further complicated by the discovery of a number of previously unknown isoforms which may be differently regulated and have different substrate specificity.
T12869	16760-16762	IN	denotes	In
T12871	16763-16766	DT	denotes	the
T12873	16767-16771	JJ	denotes	past
T12872	16772-16778	NN	denotes	decade
T12874	16778-16780	,	denotes	,
T12875	16780-16783	DT	denotes	the
T12876	16784-16789	NN	denotes	story
T12877	16790-16793	VBZ	denotes	has
T12878	16794-16798	VBN	denotes	been
T12879	16799-16806	RB	denotes	further
T12870	16807-16818	VBN	denotes	complicated
T12880	16819-16821	IN	denotes	by
T12881	16822-16825	DT	denotes	the
T12882	16826-16835	NN	denotes	discovery
T12883	16836-16838	IN	denotes	of
T12884	16839-16840	DT	denotes	a
T12885	16841-16847	NN	denotes	number
T12886	16848-16850	IN	denotes	of
T12887	16851-16861	RB	denotes	previously
T12888	16862-16869	JJ	denotes	unknown
T12889	16870-16878	NNS	denotes	isoforms
T12890	16879-16884	WDT	denotes	which
T12892	16885-16888	MD	denotes	may
T12893	16889-16891	VB	denotes	be
T12894	16892-16903	RB	denotes	differently
T12891	16904-16913	VBN	denotes	regulated
T12895	16914-16917	CC	denotes	and
T12896	16918-16922	VBP	denotes	have
T12897	16923-16932	JJ	denotes	different
T12899	16933-16942	NN	denotes	substrate
T12898	16943-16954	NN	denotes	specificity
T12900	16954-16955	.	denotes	.
T12901	16955-17076	sentence	denotes	Gene targeting in mice has provided a powerful approach to examine the physiologic roles of individual calpain isoforms.
T12902	16956-16960	NN	denotes	Gene
T12903	16961-16970	NN	denotes	targeting
T12905	16971-16973	IN	denotes	in
T12906	16974-16978	NNS	denotes	mice
T12907	16979-16982	VBZ	denotes	has
T12904	16983-16991	VBN	denotes	provided
T12908	16992-16993	DT	denotes	a
T12910	16994-17002	JJ	denotes	powerful
T12909	17003-17011	NN	denotes	approach
T12911	17012-17014	TO	denotes	to
T12912	17015-17022	VB	denotes	examine
T12913	17023-17026	DT	denotes	the
T12915	17027-17038	JJ	denotes	physiologic
T12914	17039-17044	NNS	denotes	roles
T12916	17045-17047	IN	denotes	of
T12917	17048-17058	JJ	denotes	individual
T12919	17059-17066	NN	denotes	calpain
T12918	17067-17075	NNS	denotes	isoforms
T12920	17075-17076	.	denotes	.
T12921	17076-17194	sentence	denotes	This was first used to disrupt the Capn4 gene, encoding the small regulatory subunit common to both μ- and m-calpain.
T12922	17077-17081	DT	denotes	This
T12924	17082-17085	VBD	denotes	was
T12925	17086-17091	RB	denotes	first
T12923	17092-17096	VBN	denotes	used
T12926	17097-17099	TO	denotes	to
T12927	17100-17107	VB	denotes	disrupt
T12928	17108-17111	DT	denotes	the
T12930	17112-17117	NN	denotes	Capn4
T12929	17118-17122	NN	denotes	gene
T12931	17122-17124	,	denotes	,
T12932	17124-17132	VBG	denotes	encoding
T12933	17133-17136	DT	denotes	the
T12935	17137-17142	JJ	denotes	small
T12936	17143-17153	JJ	denotes	regulatory
T12934	17154-17161	NN	denotes	subunit
T12937	17162-17168	JJ	denotes	common
T12938	17169-17171	IN	denotes	to
T12939	17172-17176	CC	denotes	both
T12941	17177-17178	NN	denotes	μ
T12942	17178-17179	HYPH	denotes	-
T12943	17180-17183	CC	denotes	and
T12944	17184-17185	NN	denotes	m
T12945	17185-17186	HYPH	denotes	-
T12940	17186-17193	NN	denotes	calpain
T12946	17193-17194	.	denotes	.
T12947	17194-17327	sentence	denotes	Two independent laboratories observed embryonic lethality in Capn4 knockout mice, albeit at different stages of development [31,32].
T12948	17195-17198	CD	denotes	Two
T12950	17199-17210	JJ	denotes	independent
T12949	17211-17223	NNS	denotes	laboratories
T12951	17224-17232	VBD	denotes	observed
T12952	17233-17242	JJ	denotes	embryonic
T12953	17243-17252	NN	denotes	lethality
T12954	17253-17255	IN	denotes	in
T12955	17256-17261	NN	denotes	Capn4
T12957	17262-17270	NN	denotes	knockout
T12956	17271-17275	NNS	denotes	mice
T12958	17275-17277	,	denotes	,
T12959	17277-17283	IN	denotes	albeit
T12960	17284-17286	IN	denotes	at
T12961	17287-17296	JJ	denotes	different
T12962	17297-17303	NNS	denotes	stages
T12963	17304-17306	IN	denotes	of
T12964	17307-17318	NN	denotes	development
T12965	17319-17320	-LRB-	denotes	[
T12967	17320-17322	CD	denotes	31
T12968	17322-17323	,	denotes	,
T12966	17323-17325	CD	denotes	32
T12969	17325-17326	-RRB-	denotes	]
T12970	17326-17327	.	denotes	.
T12971	17327-17700	sentence	denotes	These observations supported the hypothesis that the small subunit is required for both μ- and m-calpain, and furthermore suggested four possibilities regarding their requirement for embryonic development: 1) both isoforms were required; 2) μ-calpain was required; 3) m-calpain was required, or 4) μ- and m-calpain are redundant, and one or the other isoform was required.
T12972	17328-17333	DT	denotes	These
T12973	17334-17346	NNS	denotes	observations
T12974	17347-17356	VBD	denotes	supported
T12976	17357-17360	DT	denotes	the
T12977	17361-17371	NN	denotes	hypothesis
T12978	17372-17376	IN	denotes	that
T12980	17377-17380	DT	denotes	the
T12982	17381-17386	JJ	denotes	small
T12981	17387-17394	NN	denotes	subunit
T12983	17395-17397	VBZ	denotes	is
T12979	17398-17406	VBN	denotes	required
T12984	17407-17410	IN	denotes	for
T12985	17411-17415	CC	denotes	both
T12987	17416-17417	NN	denotes	μ
T12988	17417-17418	HYPH	denotes	-
T12989	17419-17422	CC	denotes	and
T12990	17423-17424	NN	denotes	m
T12991	17424-17425	HYPH	denotes	-
T12986	17425-17432	NN	denotes	calpain
T12992	17432-17434	,	denotes	,
T12993	17434-17437	CC	denotes	and
T12994	17438-17449	RB	denotes	furthermore
T12995	17450-17459	VBD	denotes	suggested
T12996	17460-17464	CD	denotes	four
T12997	17465-17478	NNS	denotes	possibilities
T12998	17479-17488	VBG	denotes	regarding
T12999	17489-17494	PRP$	denotes	their
T13000	17495-17506	NN	denotes	requirement
T13001	17507-17510	IN	denotes	for
T13002	17511-17520	JJ	denotes	embryonic
T13003	17521-17532	NN	denotes	development
T13004	17532-17534	:	denotes	:
T13005	17534-17535	LS	denotes	1
T13006	17535-17536	-RRB-	denotes	)
T13007	17537-17541	DT	denotes	both
T13008	17542-17550	NNS	denotes	isoforms
T13009	17551-17555	VBD	denotes	were
T12975	17556-17564	VBN	denotes	required
T13010	17564-17565	:	denotes	;
T13011	17566-17567	LS	denotes	2
T13013	17567-17568	-RRB-	denotes	)
T13014	17569-17570	NN	denotes	μ
T13016	17570-17571	HYPH	denotes	-
T13015	17571-17578	NN	denotes	calpain
T13017	17579-17582	VBD	denotes	was
T13012	17583-17591	VBN	denotes	required
T13018	17591-17592	:	denotes	;
T13019	17593-17594	LS	denotes	3
T13021	17594-17595	-RRB-	denotes	)
T13022	17596-17597	NN	denotes	m
T13024	17597-17598	HYPH	denotes	-
T13023	17598-17605	NN	denotes	calpain
T13025	17606-17609	VBD	denotes	was
T13020	17610-17618	VBN	denotes	required
T13026	17618-17620	,	denotes	,
T13027	17620-17622	CC	denotes	or
T13028	17623-17624	LS	denotes	4
T13030	17624-17625	-RRB-	denotes	)
T13031	17626-17627	NN	denotes	μ
T13033	17627-17628	HYPH	denotes	-
T13034	17629-17632	CC	denotes	and
T13035	17633-17634	NN	denotes	m
T13036	17634-17635	HYPH	denotes	-
T13032	17635-17642	NN	denotes	calpain
T13029	17643-17646	VBP	denotes	are
T13037	17647-17656	JJ	denotes	redundant
T13038	17656-17658	,	denotes	,
T13039	17658-17661	CC	denotes	and
T13040	17662-17665	CD	denotes	one
T13042	17666-17668	CC	denotes	or
T13043	17669-17672	DT	denotes	the
T13044	17673-17678	JJ	denotes	other
T13045	17679-17686	NN	denotes	isoform
T13046	17687-17690	VBD	denotes	was
T13041	17691-17699	VBN	denotes	required
T13047	17699-17700	.	denotes	.
T13048	17700-17931	sentence	denotes	These options were narrowed down by the subsequent observation that Capn1-/- mice, which lack the μ-calpain catalytic subunit, were healthy and fertile, although platelet aggregation and clot retraction defects were observed [33].
T13049	17701-17706	DT	denotes	These
T13050	17707-17714	NNS	denotes	options
T13052	17715-17719	VBD	denotes	were
T13051	17720-17728	VBN	denotes	narrowed
T13053	17729-17733	RP	denotes	down
T13054	17734-17736	IN	denotes	by
T13055	17737-17740	DT	denotes	the
T13057	17741-17751	JJ	denotes	subsequent
T13056	17752-17763	NN	denotes	observation
T13058	17764-17768	IN	denotes	that
T13060	17769-17774	NN	denotes	Capn1
T13062	17774-17775	SYM	denotes	-
T13063	17775-17776	HYPH	denotes	/
T13064	17776-17777	SYM	denotes	-
T13061	17778-17782	NNS	denotes	mice
T13065	17782-17784	,	denotes	,
T13066	17784-17789	WDT	denotes	which
T13067	17790-17794	VBP	denotes	lack
T13068	17795-17798	DT	denotes	the
T13070	17799-17800	NN	denotes	μ
T13072	17800-17801	HYPH	denotes	-
T13071	17801-17808	NN	denotes	calpain
T13073	17809-17818	JJ	denotes	catalytic
T13069	17819-17826	NN	denotes	subunit
T13074	17826-17828	,	denotes	,
T13059	17828-17832	VBD	denotes	were
T13075	17833-17840	JJ	denotes	healthy
T13076	17841-17844	CC	denotes	and
T13077	17845-17852	JJ	denotes	fertile
T13078	17852-17854	,	denotes	,
T13079	17854-17862	IN	denotes	although
T13081	17863-17871	NN	denotes	platelet
T13082	17872-17883	NN	denotes	aggregation
T13084	17884-17887	CC	denotes	and
T13085	17888-17892	NN	denotes	clot
T13086	17893-17903	NN	denotes	retraction
T13083	17904-17911	NNS	denotes	defects
T13087	17912-17916	VBD	denotes	were
T13080	17917-17925	VBN	denotes	observed
T13088	17926-17927	-LRB-	denotes	[
T13089	17927-17929	CD	denotes	33
T13090	17929-17930	-RRB-	denotes	]
T13091	17930-17931	.	denotes	.
T13092	17931-18141	sentence	denotes	At that point, we were left with the last two possibilities that either m-calpain was specifically required during embryogenesis, or that either isoform alone was sufficient for sustaining embryonic viability.
T13093	17932-17934	IN	denotes	At
T13095	17935-17939	DT	denotes	that
T13096	17940-17945	NN	denotes	point
T13097	17945-17947	,	denotes	,
T13098	17947-17949	PRP	denotes	we
T13099	17950-17954	VBD	denotes	were
T13094	17955-17959	VBN	denotes	left
T13100	17960-17964	IN	denotes	with
T13101	17965-17968	DT	denotes	the
T13103	17969-17973	JJ	denotes	last
T13104	17974-17977	CD	denotes	two
T13102	17978-17991	NNS	denotes	possibilities
T13105	17992-17996	IN	denotes	that
T13107	17997-18003	DT	denotes	either
T13109	18004-18005	NN	denotes	m
T13110	18005-18006	HYPH	denotes	-
T13108	18006-18013	NN	denotes	calpain
T13111	18014-18017	VBD	denotes	was
T13112	18018-18030	RB	denotes	specifically
T13106	18031-18039	VBN	denotes	required
T13113	18040-18046	IN	denotes	during
T13114	18047-18060	NN	denotes	embryogenesis
T13115	18060-18062	,	denotes	,
T13116	18062-18064	CC	denotes	or
T13117	18065-18069	IN	denotes	that
T13119	18070-18076	DT	denotes	either
T13120	18077-18084	NN	denotes	isoform
T13121	18085-18090	RB	denotes	alone
T13118	18091-18094	VBD	denotes	was
T13122	18095-18105	JJ	denotes	sufficient
T13123	18106-18109	IN	denotes	for
T13124	18110-18120	VBG	denotes	sustaining
T13125	18121-18130	JJ	denotes	embryonic
T13126	18131-18140	NN	denotes	viability
T13127	18140-18141	.	denotes	.
T13128	18141-18269	sentence	denotes	We report here that Capn2-/- mice, which lack the m-calpain catalytic subunit, die at the preimplantation stage of development.
T13129	18142-18144	PRP	denotes	We
T13130	18145-18151	VBP	denotes	report
T13131	18152-18156	RB	denotes	here
T13132	18157-18161	IN	denotes	that
T13134	18162-18167	NN	denotes	Capn2
T13136	18167-18168	SYM	denotes	-
T13137	18168-18169	HYPH	denotes	/
T13138	18169-18170	SYM	denotes	-
T13135	18171-18175	NNS	denotes	mice
T13139	18175-18177	,	denotes	,
T13140	18177-18182	WDT	denotes	which
T13141	18183-18187	VBP	denotes	lack
T13142	18188-18191	DT	denotes	the
T13144	18192-18193	NN	denotes	m
T13146	18193-18194	HYPH	denotes	-
T13145	18194-18201	NN	denotes	calpain
T13147	18202-18211	JJ	denotes	catalytic
T13143	18212-18219	NN	denotes	subunit
T13148	18219-18221	,	denotes	,
T13133	18221-18224	VBP	denotes	die
T13149	18225-18227	IN	denotes	at
T13150	18228-18231	DT	denotes	the
T13152	18232-18247	NN	denotes	preimplantation
T13151	18248-18253	NN	denotes	stage
T13153	18254-18256	IN	denotes	of
T13154	18257-18268	NN	denotes	development
T13155	18268-18269	.	denotes	.
T13156	18269-18374	sentence	denotes	This observation allows us to now conclude that m-calpain is specifically required during embryogenesis.
T13157	18270-18274	DT	denotes	This
T13158	18275-18286	NN	denotes	observation
T13159	18287-18293	VBZ	denotes	allows
T13160	18294-18296	PRP	denotes	us
T13162	18297-18299	TO	denotes	to
T13163	18300-18303	RB	denotes	now
T13161	18304-18312	VB	denotes	conclude
T13164	18313-18317	IN	denotes	that
T13166	18318-18319	NN	denotes	m
T13168	18319-18320	HYPH	denotes	-
T13167	18320-18327	NN	denotes	calpain
T13169	18328-18330	VBZ	denotes	is
T13170	18331-18343	RB	denotes	specifically
T13165	18344-18352	VBN	denotes	required
T13171	18353-18359	IN	denotes	during
T13172	18360-18373	NN	denotes	embryogenesis
T13173	18373-18374	.	denotes	.
T13174	18374-18640	sentence	denotes	Since homozygous disruption of the Capn4 gene was also expected to abolish m-calpain activity, this result is in agreement with the phenotype presented by one of the two Capn4 targeted lines in which preimplantation lethality of null embryos was also observed [32].
T13175	18375-18380	IN	denotes	Since
T13177	18381-18391	JJ	denotes	homozygous
T13178	18392-18402	NN	denotes	disruption
T13179	18403-18405	IN	denotes	of
T13180	18406-18409	DT	denotes	the
T13182	18410-18415	NN	denotes	Capn4
T13181	18416-18420	NN	denotes	gene
T13183	18421-18424	VBD	denotes	was
T13184	18425-18429	RB	denotes	also
T13176	18430-18438	VBN	denotes	expected
T13186	18439-18441	TO	denotes	to
T13187	18442-18449	VB	denotes	abolish
T13188	18450-18451	NN	denotes	m
T13190	18451-18452	HYPH	denotes	-
T13189	18452-18459	NN	denotes	calpain
T13191	18460-18468	NN	denotes	activity
T13192	18468-18470	,	denotes	,
T13193	18470-18474	DT	denotes	this
T13194	18475-18481	NN	denotes	result
T13185	18482-18484	VBZ	denotes	is
T13195	18485-18487	IN	denotes	in
T13196	18488-18497	NN	denotes	agreement
T13197	18498-18502	IN	denotes	with
T13198	18503-18506	DT	denotes	the
T13199	18507-18516	NN	denotes	phenotype
T13200	18517-18526	VBN	denotes	presented
T13201	18527-18529	IN	denotes	by
T13202	18530-18533	CD	denotes	one
T13203	18534-18536	IN	denotes	of
T13204	18537-18540	DT	denotes	the
T13206	18541-18544	CD	denotes	two
T13207	18545-18550	NN	denotes	Capn4
T13208	18551-18559	VBN	denotes	targeted
T13205	18560-18565	NNS	denotes	lines
T13209	18566-18568	IN	denotes	in
T13211	18569-18574	WDT	denotes	which
T13212	18575-18590	NN	denotes	preimplantation
T13213	18591-18600	NN	denotes	lethality
T13214	18601-18603	IN	denotes	of
T13215	18604-18608	JJ	denotes	null
T13216	18609-18616	NNS	denotes	embryos
T13217	18617-18620	VBD	denotes	was
T13218	18621-18625	RB	denotes	also
T13210	18626-18634	VBN	denotes	observed
T13219	18635-18636	-LRB-	denotes	[
T13220	18636-18638	CD	denotes	32
T13221	18638-18639	-RRB-	denotes	]
T13222	18639-18640	.	denotes	.
T13223	18640-18793	sentence	denotes	The survival of Capn4-/- embryos from the original targeted line reported by Arthur and colleagues to mid-gestation is more difficult to reconcile [31].
T13224	18641-18644	DT	denotes	The
T13225	18645-18653	NN	denotes	survival
T13227	18654-18656	IN	denotes	of
T13228	18657-18662	NN	denotes	Capn4
T13230	18662-18663	SYM	denotes	-
T13231	18663-18664	HYPH	denotes	/
T13232	18664-18665	SYM	denotes	-
T13229	18666-18673	NNS	denotes	embryos
T13233	18674-18678	IN	denotes	from
T13234	18679-18682	DT	denotes	the
T13236	18683-18691	JJ	denotes	original
T13237	18692-18700	VBN	denotes	targeted
T13235	18701-18705	NN	denotes	line
T13238	18706-18714	VBN	denotes	reported
T13239	18715-18717	IN	denotes	by
T13240	18718-18724	NNP	denotes	Arthur
T13241	18725-18728	CC	denotes	and
T13242	18729-18739	NNS	denotes	colleagues
T13243	18740-18742	IN	denotes	to
T13244	18743-18756	NN	denotes	mid-gestation
T13226	18757-18759	VBZ	denotes	is
T13245	18760-18764	RBR	denotes	more
T13246	18765-18774	JJ	denotes	difficult
T13247	18775-18777	TO	denotes	to
T13248	18778-18787	VB	denotes	reconcile
T13249	18788-18789	-LRB-	denotes	[
T13250	18789-18791	CD	denotes	31
T13251	18791-18792	-RRB-	denotes	]
T13252	18792-18793	.	denotes	.
T13253	18793-18902	sentence	denotes	In retrospect, it seems likely that the latter line represents a hypomorphic state, rather than a true null.
T13254	18794-18796	IN	denotes	In
T13256	18797-18807	NN	denotes	retrospect
T13257	18807-18809	,	denotes	,
T13258	18809-18811	PRP	denotes	it
T13255	18812-18817	VBZ	denotes	seems
T13259	18818-18824	JJ	denotes	likely
T13260	18825-18829	IN	denotes	that
T13262	18830-18833	DT	denotes	the
T13264	18834-18840	JJ	denotes	latter
T13263	18841-18845	NN	denotes	line
T13261	18846-18856	VBZ	denotes	represents
T13265	18857-18858	DT	denotes	a
T13267	18859-18870	JJ	denotes	hypomorphic
T13266	18871-18876	NN	denotes	state
T13268	18876-18878	,	denotes	,
T13269	18878-18884	RB	denotes	rather
T13270	18885-18889	IN	denotes	than
T13271	18890-18891	DT	denotes	a
T13273	18892-18896	JJ	denotes	true
T13272	18897-18901	NN	denotes	null
T13274	18901-18902	.	denotes	.
T13275	18902-19138	sentence	denotes	The Capn4 targeting strategy employed by Arthur and colleagues involved disrupting the C-terminus of the calpain small subunit by insertion of the PGK-Neo cassette into the middle of exon 9, which caused truncation of the protein [31].
T13276	18903-18906	DT	denotes	The
T13278	18907-18912	NN	denotes	Capn4
T13279	18913-18922	NN	denotes	targeting
T13277	18923-18931	NN	denotes	strategy
T13281	18932-18940	VBN	denotes	employed
T13282	18941-18943	IN	denotes	by
T13283	18944-18950	NNP	denotes	Arthur
T13284	18951-18954	CC	denotes	and
T13285	18955-18965	NNS	denotes	colleagues
T13280	18966-18974	VBN	denotes	involved
T13286	18975-18985	VBG	denotes	disrupting
T13287	18986-18989	DT	denotes	the
T13289	18990-18991	NN	denotes	C
T13290	18991-18992	HYPH	denotes	-
T13288	18992-19000	NN	denotes	terminus
T13291	19001-19003	IN	denotes	of
T13292	19004-19007	DT	denotes	the
T13294	19008-19015	NN	denotes	calpain
T13295	19016-19021	JJ	denotes	small
T13293	19022-19029	NN	denotes	subunit
T13296	19030-19032	IN	denotes	by
T13297	19033-19042	NN	denotes	insertion
T13298	19043-19045	IN	denotes	of
T13299	19046-19049	DT	denotes	the
T13301	19050-19053	NN	denotes	PGK
T13303	19053-19054	HYPH	denotes	-
T13302	19054-19057	NN	denotes	Neo
T13300	19058-19066	NN	denotes	cassette
T13304	19067-19071	IN	denotes	into
T13305	19072-19075	DT	denotes	the
T13306	19076-19082	NN	denotes	middle
T13307	19083-19085	IN	denotes	of
T13308	19086-19090	NN	denotes	exon
T13309	19091-19092	CD	denotes	9
T13310	19092-19094	,	denotes	,
T13311	19094-19099	WDT	denotes	which
T13312	19100-19106	VBD	denotes	caused
T13313	19107-19117	NN	denotes	truncation
T13314	19118-19120	IN	denotes	of
T13315	19121-19124	DT	denotes	the
T13316	19125-19132	NN	denotes	protein
T13317	19133-19134	-LRB-	denotes	[
T13318	19134-19136	CD	denotes	31
T13319	19136-19137	-RRB-	denotes	]
T13320	19137-19138	.	denotes	.
T13321	19138-19382	sentence	denotes	This strategy was based upon previous structure/function studies showing that excision of the C-terminal 25 amino acid residues of the small subunit abolished calpain activity when co-expressed with the m-calpain large subunit in E. coli [34].
T13322	19139-19143	DT	denotes	This
T13323	19144-19152	NN	denotes	strategy
T13325	19153-19156	VBD	denotes	was
T13324	19157-19162	VBN	denotes	based
T13326	19163-19167	IN	denotes	upon
T13327	19168-19176	JJ	denotes	previous
T13329	19177-19186	NN	denotes	structure
T13331	19186-19187	HYPH	denotes	/
T13330	19187-19195	NN	denotes	function
T13328	19196-19203	NNS	denotes	studies
T13332	19204-19211	VBG	denotes	showing
T13333	19212-19216	IN	denotes	that
T13335	19217-19225	NN	denotes	excision
T13336	19226-19228	IN	denotes	of
T13337	19229-19232	DT	denotes	the
T13339	19233-19234	NN	denotes	C
T13341	19234-19235	HYPH	denotes	-
T13340	19235-19243	JJ	denotes	terminal
T13342	19244-19246	CD	denotes	25
T13343	19247-19252	NN	denotes	amino
T13338	19253-19257	NN	denotes	acid
T13344	19258-19266	VBZ	denotes	residues
T13345	19267-19269	IN	denotes	of
T13346	19270-19273	DT	denotes	the
T13348	19274-19279	JJ	denotes	small
T13347	19280-19287	NN	denotes	subunit
T13334	19288-19297	VBD	denotes	abolished
T13349	19298-19305	NN	denotes	calpain
T13350	19306-19314	NN	denotes	activity
T13351	19315-19319	WRB	denotes	when
T13352	19320-19332	VBN	denotes	co-expressed
T13353	19333-19337	IN	denotes	with
T13354	19338-19341	DT	denotes	the
T13356	19342-19343	NN	denotes	m
T13358	19343-19344	HYPH	denotes	-
T13357	19344-19351	NN	denotes	calpain
T13359	19352-19357	JJ	denotes	large
T13355	19358-19365	NN	denotes	subunit
T13360	19366-19368	IN	denotes	in
T13361	19369-19371	NNP	denotes	E.
T13362	19372-19376	NNP	denotes	coli
T13363	19377-19378	-LRB-	denotes	[
T13364	19378-19380	CD	denotes	34
T13365	19380-19381	-RRB-	denotes	]
T13366	19381-19382	.	denotes	.
T13367	19382-19528	sentence	denotes	In contrast, the Capn4 targeting strategy employed by Zimmerman and colleagues involved a much more extensive deletion of exons 4 through 8 [32].
T13368	19383-19385	IN	denotes	In
T13370	19386-19394	NN	denotes	contrast
T13371	19394-19396	,	denotes	,
T13372	19396-19399	DT	denotes	the
T13374	19400-19405	NN	denotes	Capn4
T13375	19406-19415	VBG	denotes	targeting
T13373	19416-19424	NN	denotes	strategy
T13376	19425-19433	VBN	denotes	employed
T13377	19434-19436	IN	denotes	by
T13378	19437-19446	NNP	denotes	Zimmerman
T13379	19447-19450	CC	denotes	and
T13380	19451-19461	NNS	denotes	colleagues
T13369	19462-19470	VBD	denotes	involved
T13381	19471-19472	DT	denotes	a
T13383	19473-19477	RB	denotes	much
T13384	19478-19482	RBR	denotes	more
T13385	19483-19492	JJ	denotes	extensive
T13382	19493-19501	NN	denotes	deletion
T13386	19502-19504	IN	denotes	of
T13387	19505-19510	NNS	denotes	exons
T13388	19511-19512	CD	denotes	4
T13389	19513-19520	IN	denotes	through
T13390	19521-19522	CD	denotes	8
T13391	19523-19524	-LRB-	denotes	[
T13392	19524-19526	CD	denotes	32
T13393	19526-19527	-RRB-	denotes	]
T13394	19527-19528	.	denotes	.
T13395	19528-19682	sentence	denotes	It now seems probable that the difference in the time of lethality of these two targeted Capn4 lines can be explained by different extents of disruption.
T13396	19529-19531	PRP	denotes	It
T13398	19532-19535	RB	denotes	now
T13397	19536-19541	VBZ	denotes	seems
T13399	19542-19550	JJ	denotes	probable
T13400	19551-19555	IN	denotes	that
T13402	19556-19559	DT	denotes	the
T13403	19560-19570	NN	denotes	difference
T13404	19571-19573	IN	denotes	in
T13405	19574-19577	DT	denotes	the
T13406	19578-19582	NN	denotes	time
T13407	19583-19585	IN	denotes	of
T13408	19586-19595	NN	denotes	lethality
T13409	19596-19598	IN	denotes	of
T13410	19599-19604	DT	denotes	these
T13412	19605-19608	CD	denotes	two
T13413	19609-19617	VBN	denotes	targeted
T13414	19618-19623	NN	denotes	Capn4
T13411	19624-19629	NNS	denotes	lines
T13415	19630-19633	MD	denotes	can
T13416	19634-19636	VB	denotes	be
T13401	19637-19646	VBN	denotes	explained
T13417	19647-19649	IN	denotes	by
T13418	19650-19659	JJ	denotes	different
T13419	19660-19667	NNS	denotes	extents
T13420	19668-19670	IN	denotes	of
T13421	19671-19681	NN	denotes	disruption
T13422	19681-19682	.	denotes	.
T13423	19682-19802	sentence	denotes	The Zimmerman allele probably represents a true null genotype while the Arthur allele is likely a hypomorphic mutation.
T13424	19683-19686	DT	denotes	The
T13426	19687-19696	NNP	denotes	Zimmerman
T13425	19697-19703	NN	denotes	allele
T13428	19704-19712	RB	denotes	probably
T13427	19713-19723	VBZ	denotes	represents
T13429	19724-19725	DT	denotes	a
T13431	19726-19730	JJ	denotes	true
T13432	19731-19735	JJ	denotes	null
T13430	19736-19744	NN	denotes	genotype
T13433	19745-19750	IN	denotes	while
T13435	19751-19754	DT	denotes	the
T13437	19755-19761	NNP	denotes	Arthur
T13436	19762-19768	NN	denotes	allele
T13434	19769-19771	VBZ	denotes	is
T13438	19772-19778	RB	denotes	likely
T13439	19779-19780	DT	denotes	a
T13441	19781-19792	JJ	denotes	hypomorphic
T13440	19793-19801	NN	denotes	mutation
T13442	19801-19802	.	denotes	.
T13443	19802-19907	sentence	denotes	Alternate targeting strategies have been shown to yield different phenotypes in gene disruption studies.
T13444	19803-19812	JJ	denotes	Alternate
T13446	19813-19822	NN	denotes	targeting
T13445	19823-19833	NNS	denotes	strategies
T13448	19834-19838	VBP	denotes	have
T13449	19839-19843	VBN	denotes	been
T13447	19844-19849	VBN	denotes	shown
T13450	19850-19852	TO	denotes	to
T13451	19853-19858	VB	denotes	yield
T13452	19859-19868	JJ	denotes	different
T13453	19869-19879	NNS	denotes	phenotypes
T13454	19880-19882	IN	denotes	in
T13455	19883-19887	NN	denotes	gene
T13456	19888-19898	NN	denotes	disruption
T13457	19899-19906	NNS	denotes	studies
T13458	19906-19907	.	denotes	.
T13459	19907-20031	sentence	denotes	For example, three different targeting strategies were independently used to disrupt the mixed lineage leukemia (Mll) gene.
T13460	19908-19911	IN	denotes	For
T13462	19912-19919	NN	denotes	example
T13463	19919-19921	,	denotes	,
T13464	19921-19926	CD	denotes	three
T13466	19927-19936	JJ	denotes	different
T13467	19937-19946	NN	denotes	targeting
T13465	19947-19957	NNS	denotes	strategies
T13468	19958-19962	VBD	denotes	were
T13469	19963-19976	RB	denotes	independently
T13461	19977-19981	VBN	denotes	used
T13470	19982-19984	TO	denotes	to
T13471	19985-19992	VB	denotes	disrupt
T13472	19993-19996	DT	denotes	the
T13474	19997-20002	JJ	denotes	mixed
T13476	20003-20010	NN	denotes	lineage
T13475	20011-20019	NN	denotes	leukemia
T13477	20020-20021	-LRB-	denotes	(
T13478	20021-20024	NN	denotes	Mll
T13479	20024-20025	-RRB-	denotes	)
T13473	20026-20030	NN	denotes	gene
T13480	20030-20031	.	denotes	.
T13481	20031-20159	sentence	denotes	In all three studies, homozygous null embryos perished during embryogenesis, but at different stages (E0.5, E10.5, E14.5) [35].
T13482	20032-20034	IN	denotes	In
T13484	20035-20038	DT	denotes	all
T13486	20039-20044	CD	denotes	three
T13485	20045-20052	NNS	denotes	studies
T13487	20052-20054	,	denotes	,
T13488	20054-20064	JJ	denotes	homozygous
T13489	20065-20069	JJ	denotes	null
T13490	20070-20077	NNS	denotes	embryos
T13483	20078-20086	VBN	denotes	perished
T13491	20087-20093	IN	denotes	during
T13492	20094-20107	NN	denotes	embryogenesis
T13493	20107-20109	,	denotes	,
T13494	20109-20112	CC	denotes	but
T13495	20113-20115	IN	denotes	at
T13496	20116-20125	JJ	denotes	different
T13497	20126-20132	NNS	denotes	stages
T13498	20133-20134	-LRB-	denotes	(
T13500	20134-20138	NN	denotes	E0.5
T13501	20138-20140	,	denotes	,
T13502	20140-20145	NN	denotes	E10.5
T13503	20145-20147	,	denotes	,
T13499	20147-20152	NN	denotes	E14.5
T13504	20152-20153	-RRB-	denotes	)
T13505	20154-20155	-LRB-	denotes	[
T13506	20155-20157	CD	denotes	35
T13507	20157-20158	-RRB-	denotes	]
T13508	20158-20159	.	denotes	.
T13509	20159-20302	sentence	denotes	The variation in phenotype was attributed to the differences in degree of function of the truncated proteins produced from the mutant alleles.
T13510	20160-20163	DT	denotes	The
T13511	20164-20173	NN	denotes	variation
T13513	20174-20176	IN	denotes	in
T13514	20177-20186	NN	denotes	phenotype
T13515	20187-20190	VBD	denotes	was
T13512	20191-20201	VBN	denotes	attributed
T13516	20202-20204	IN	denotes	to
T13517	20205-20208	DT	denotes	the
T13518	20209-20220	NNS	denotes	differences
T13519	20221-20223	IN	denotes	in
T13520	20224-20230	NN	denotes	degree
T13521	20231-20233	IN	denotes	of
T13522	20234-20242	NN	denotes	function
T13523	20243-20245	IN	denotes	of
T13524	20246-20249	DT	denotes	the
T13526	20250-20259	VBN	denotes	truncated
T13525	20260-20268	NN	denotes	proteins
T13527	20269-20277	VBN	denotes	produced
T13528	20278-20282	IN	denotes	from
T13529	20283-20286	DT	denotes	the
T13531	20287-20293	NN	denotes	mutant
T13530	20294-20301	NNS	denotes	alleles
T13532	20301-20302	.	denotes	.
T13533	20302-20371	sentence	denotes	A similar effect might be at work in the two Capn4 transgenic lines.
T13534	20303-20304	DT	denotes	A
T13536	20305-20312	JJ	denotes	similar
T13535	20313-20319	NN	denotes	effect
T13538	20320-20325	MD	denotes	might
T13537	20326-20328	VB	denotes	be
T13539	20329-20331	IN	denotes	at
T13540	20332-20336	NN	denotes	work
T13541	20337-20339	IN	denotes	in
T13542	20340-20343	DT	denotes	the
T13544	20344-20347	CD	denotes	two
T13545	20348-20353	NN	denotes	Capn4
T13546	20354-20364	JJ	denotes	transgenic
T13543	20365-20370	NNS	denotes	lines
T13547	20370-20371	.	denotes	.
T13548	20371-20471	sentence	denotes	Efforts were made to detect transcripts or calpain activities derived from the Arthur Capn4 allele.
T13549	20372-20379	NNS	denotes	Efforts
T13551	20380-20384	VBD	denotes	were
T13550	20385-20389	VBN	denotes	made
T13552	20390-20392	TO	denotes	to
T13553	20393-20399	VB	denotes	detect
T13554	20400-20411	NNS	denotes	transcripts
T13555	20412-20414	CC	denotes	or
T13556	20415-20422	NN	denotes	calpain
T13557	20423-20433	NNS	denotes	activities
T13558	20434-20441	VBN	denotes	derived
T13559	20442-20446	IN	denotes	from
T13560	20447-20450	DT	denotes	the
T13562	20451-20457	NNP	denotes	Arthur
T13563	20458-20463	NN	denotes	Capn4
T13561	20464-20470	NN	denotes	allele
T13564	20470-20471	.	denotes	.
T13565	20471-20669	sentence	denotes	This allele gave rise to multiple mRNA species, detectable by RT-PCR, reading through from the first half of exon 9 to at least two different cryptic splice sites in the PGK promoter sequence [31].
T13566	20472-20476	DT	denotes	This
T13567	20477-20483	NN	denotes	allele
T13568	20484-20488	VBD	denotes	gave
T13569	20489-20493	NN	denotes	rise
T13570	20494-20496	IN	denotes	to
T13571	20497-20505	JJ	denotes	multiple
T13573	20506-20510	NN	denotes	mRNA
T13572	20511-20518	NNS	denotes	species
T13574	20518-20520	,	denotes	,
T13575	20520-20530	JJ	denotes	detectable
T13576	20531-20533	IN	denotes	by
T13577	20534-20536	NN	denotes	RT
T13579	20536-20537	HYPH	denotes	-
T13578	20537-20540	NN	denotes	PCR
T13580	20540-20542	,	denotes	,
T13581	20542-20549	VBG	denotes	reading
T13582	20550-20557	RB	denotes	through
T13583	20558-20562	IN	denotes	from
T13584	20563-20566	DT	denotes	the
T13586	20567-20572	JJ	denotes	first
T13585	20573-20577	NN	denotes	half
T13587	20578-20580	IN	denotes	of
T13588	20581-20585	NN	denotes	exon
T13589	20586-20587	CD	denotes	9
T13590	20588-20590	IN	denotes	to
T13591	20591-20593	RB	denotes	at
T13593	20594-20599	RBS	denotes	least
T13592	20600-20603	CD	denotes	two
T13595	20604-20613	JJ	denotes	different
T13596	20614-20621	JJ	denotes	cryptic
T13597	20622-20628	NN	denotes	splice
T13594	20629-20634	NNS	denotes	sites
T13598	20635-20637	IN	denotes	in
T13599	20638-20641	DT	denotes	the
T13601	20642-20645	NN	denotes	PGK
T13602	20646-20654	NN	denotes	promoter
T13600	20655-20663	NN	denotes	sequence
T13603	20664-20665	-LRB-	denotes	[
T13604	20665-20667	CD	denotes	31
T13605	20667-20668	-RRB-	denotes	]
T13606	20668-20669	.	denotes	.
T13607	20669-20852	sentence	denotes	These transcripts could give rise to defective calpain small subunits with 10–30 inappropriate C-terminal acids, which might be sufficient to support a low level of calpain activity.
T13608	20670-20675	DT	denotes	These
T13609	20676-20687	NNS	denotes	transcripts
T13611	20688-20693	MD	denotes	could
T13610	20694-20698	VB	denotes	give
T13612	20699-20703	NN	denotes	rise
T13613	20704-20706	IN	denotes	to
T13614	20707-20716	JJ	denotes	defective
T13616	20717-20724	NN	denotes	calpain
T13617	20725-20730	JJ	denotes	small
T13615	20731-20739	NNS	denotes	subunits
T13618	20740-20744	IN	denotes	with
T13619	20745-20747	CD	denotes	10
T13621	20747-20748	SYM	denotes	–
T13620	20748-20750	CD	denotes	30
T13623	20751-20764	JJ	denotes	inappropriate
T13624	20765-20766	NN	denotes	C
T13626	20766-20767	HYPH	denotes	-
T13625	20767-20775	JJ	denotes	terminal
T13622	20776-20781	NNS	denotes	acids
T13627	20781-20783	,	denotes	,
T13628	20783-20788	WDT	denotes	which
T13630	20789-20794	MD	denotes	might
T13629	20795-20797	VB	denotes	be
T13631	20798-20808	JJ	denotes	sufficient
T13632	20809-20811	TO	denotes	to
T13633	20812-20819	VB	denotes	support
T13634	20820-20821	DT	denotes	a
T13636	20822-20825	JJ	denotes	low
T13635	20826-20831	NN	denotes	level
T13637	20832-20834	IN	denotes	of
T13638	20835-20842	NN	denotes	calpain
T13639	20843-20851	NN	denotes	activity
T13640	20851-20852	.	denotes	.
T13641	20852-21433	sentence	denotes	However, expression of calpains with these modified small subunits did not give rise to any detectable calpain activity when expressed in E. coli, although their functionality in mammalian cells has yet to be determined (J.S. Elce, unpublished work) It has also been suggested that calpain large subunits alone might provide some activity in eukaryotic cells, although the Zimmerman et al. knockout appears to exclude that possibility, and no calpain activity was observed in our hands when calpain large subunits were expressed alone either in E. coli or in mammalian cells [36].
T13642	20853-20860	RB	denotes	However
T13644	20860-20862	,	denotes	,
T13645	20862-20872	NN	denotes	expression
T13646	20873-20875	IN	denotes	of
T13647	20876-20884	NNS	denotes	calpains
T13648	20885-20889	IN	denotes	with
T13649	20890-20895	DT	denotes	these
T13651	20896-20904	VBN	denotes	modified
T13652	20905-20910	JJ	denotes	small
T13650	20911-20919	NNS	denotes	subunits
T13653	20920-20923	VBD	denotes	did
T13654	20924-20927	RB	denotes	not
T13643	20928-20932	VB	denotes	give
T13656	20933-20937	NN	denotes	rise
T13657	20938-20940	IN	denotes	to
T13658	20941-20944	DT	denotes	any
T13660	20945-20955	JJ	denotes	detectable
T13661	20956-20963	NN	denotes	calpain
T13659	20964-20972	NN	denotes	activity
T13662	20973-20977	WRB	denotes	when
T13663	20978-20987	VBN	denotes	expressed
T13664	20988-20990	IN	denotes	in
T13665	20991-20993	NNP	denotes	E.
T13666	20994-20998	NNP	denotes	coli
T13667	20998-21000	,	denotes	,
T13668	21000-21008	IN	denotes	although
T13670	21009-21014	PRP$	denotes	their
T13671	21015-21028	NN	denotes	functionality
T13672	21029-21031	IN	denotes	in
T13673	21032-21041	JJ	denotes	mammalian
T13674	21042-21047	NNS	denotes	cells
T13669	21048-21051	VBZ	denotes	has
T13675	21052-21055	RB	denotes	yet
T13676	21056-21058	TO	denotes	to
T13678	21059-21061	VB	denotes	be
T13677	21062-21072	VBN	denotes	determined
T13679	21073-21074	-LRB-	denotes	(
T13680	21074-21078	NNP	denotes	J.S.
T13681	21079-21083	NNP	denotes	Elce
T13682	21083-21085	,	denotes	,
T13683	21085-21096	JJ	denotes	unpublished
T13684	21097-21101	NN	denotes	work
T13685	21101-21102	-RRB-	denotes	)
T13686	21103-21105	PRP	denotes	It
T13687	21106-21109	VBZ	denotes	has
T13688	21110-21114	RB	denotes	also
T13689	21115-21119	VBN	denotes	been
T13655	21120-21129	VBN	denotes	suggested
T13690	21130-21134	IN	denotes	that
T13692	21135-21142	NN	denotes	calpain
T13694	21143-21148	JJ	denotes	large
T13693	21149-21157	NNS	denotes	subunits
T13695	21158-21163	RB	denotes	alone
T13696	21164-21169	MD	denotes	might
T13691	21170-21177	VB	denotes	provide
T13697	21178-21182	DT	denotes	some
T13698	21183-21191	NN	denotes	activity
T13699	21192-21194	IN	denotes	in
T13700	21195-21205	JJ	denotes	eukaryotic
T13701	21206-21211	NNS	denotes	cells
T13702	21211-21213	,	denotes	,
T13703	21213-21221	IN	denotes	although
T13705	21222-21225	DT	denotes	the
T13707	21226-21235	NNP	denotes	Zimmerman
T13708	21236-21238	FW	denotes	et
T13709	21239-21242	FW	denotes	al.
T13706	21243-21251	NN	denotes	knockout
T13704	21252-21259	VBZ	denotes	appears
T13710	21260-21262	TO	denotes	to
T13711	21263-21270	VB	denotes	exclude
T13712	21271-21275	DT	denotes	that
T13713	21276-21287	NN	denotes	possibility
T13714	21287-21289	,	denotes	,
T13715	21289-21292	CC	denotes	and
T13716	21293-21295	DT	denotes	no
T13718	21296-21303	NN	denotes	calpain
T13717	21304-21312	NN	denotes	activity
T13720	21313-21316	VBD	denotes	was
T13719	21317-21325	VBN	denotes	observed
T13721	21326-21328	IN	denotes	in
T13722	21329-21332	PRP$	denotes	our
T13723	21333-21338	NNS	denotes	hands
T13724	21339-21343	WRB	denotes	when
T13726	21344-21351	NN	denotes	calpain
T13728	21352-21357	JJ	denotes	large
T13727	21358-21366	NNS	denotes	subunits
T13729	21367-21371	VBD	denotes	were
T13725	21372-21381	VBN	denotes	expressed
T13730	21382-21387	RB	denotes	alone
T13731	21388-21394	CC	denotes	either
T13732	21395-21397	IN	denotes	in
T13733	21398-21400	NNP	denotes	E.
T13734	21401-21405	NNP	denotes	coli
T13735	21406-21408	CC	denotes	or
T13736	21409-21411	IN	denotes	in
T13737	21412-21421	JJ	denotes	mammalian
T13738	21422-21427	NNS	denotes	cells
T13739	21428-21429	-LRB-	denotes	[
T13740	21429-21431	CD	denotes	36
T13741	21431-21432	-RRB-	denotes	]
T13742	21432-21433	.	denotes	.
T13743	21433-21685	sentence	denotes	The different timing of lethality in the Capn4 knockouts might also be a consequence of the different genetic backgrounds of the two transgenic lines, which has been observed to influence the phenotype of transgenic mice on a number of occasions [37].
T13744	21434-21437	DT	denotes	The
T13746	21438-21447	JJ	denotes	different
T13745	21448-21454	NN	denotes	timing
T13748	21455-21457	IN	denotes	of
T13749	21458-21467	NN	denotes	lethality
T13750	21468-21470	IN	denotes	in
T13751	21471-21474	DT	denotes	the
T13753	21475-21480	NN	denotes	Capn4
T13752	21481-21490	NNS	denotes	knockouts
T13754	21491-21496	MD	denotes	might
T13755	21497-21501	RB	denotes	also
T13747	21502-21504	VB	denotes	be
T13756	21505-21506	DT	denotes	a
T13757	21507-21518	NN	denotes	consequence
T13758	21519-21521	IN	denotes	of
T13759	21522-21525	DT	denotes	the
T13761	21526-21535	JJ	denotes	different
T13762	21536-21543	JJ	denotes	genetic
T13760	21544-21555	NNS	denotes	backgrounds
T13763	21556-21558	IN	denotes	of
T13764	21559-21562	DT	denotes	the
T13766	21563-21566	CD	denotes	two
T13767	21567-21577	JJ	denotes	transgenic
T13765	21578-21583	NNS	denotes	lines
T13768	21583-21585	,	denotes	,
T13769	21585-21590	WDT	denotes	which
T13771	21591-21594	VBZ	denotes	has
T13772	21595-21599	VBN	denotes	been
T13770	21600-21608	VBN	denotes	observed
T13773	21609-21611	TO	denotes	to
T13774	21612-21621	VB	denotes	influence
T13775	21622-21625	DT	denotes	the
T13776	21626-21635	NN	denotes	phenotype
T13777	21636-21638	IN	denotes	of
T13778	21639-21649	JJ	denotes	transgenic
T13779	21650-21654	NNS	denotes	mice
T13780	21655-21657	IN	denotes	on
T13781	21658-21659	DT	denotes	a
T13782	21660-21666	NN	denotes	number
T13783	21667-21669	IN	denotes	of
T13784	21670-21679	NNS	denotes	occasions
T13785	21680-21681	-LRB-	denotes	[
T13786	21681-21683	CD	denotes	37
T13787	21683-21684	-RRB-	denotes	]
T13788	21684-21685	.	denotes	.
T13789	21685-21831	sentence	denotes	One of the enduring questions in calpain research has been whether the two ubiquitous isoforms, μ- and m-calpain, possess distinct in vivo roles.
T13790	21686-21689	CD	denotes	One
T13792	21690-21692	IN	denotes	of
T13793	21693-21696	DT	denotes	the
T13795	21697-21705	VBG	denotes	enduring
T13794	21706-21715	NNS	denotes	questions
T13796	21716-21718	IN	denotes	in
T13797	21719-21726	NN	denotes	calpain
T13798	21727-21735	NN	denotes	research
T13799	21736-21739	VBZ	denotes	has
T13791	21740-21744	VBN	denotes	been
T13800	21745-21752	IN	denotes	whether
T13802	21753-21756	DT	denotes	the
T13804	21757-21760	CD	denotes	two
T13805	21761-21771	JJ	denotes	ubiquitous
T13803	21772-21780	NNS	denotes	isoforms
T13806	21780-21782	,	denotes	,
T13807	21782-21783	NN	denotes	μ
T13809	21783-21784	HYPH	denotes	-
T13810	21785-21788	CC	denotes	and
T13811	21789-21790	NN	denotes	m
T13812	21790-21791	HYPH	denotes	-
T13808	21791-21798	NN	denotes	calpain
T13813	21798-21800	,	denotes	,
T13801	21800-21807	VBP	denotes	possess
T13814	21808-21816	JJ	denotes	distinct
T13816	21817-21819	FW	denotes	in
T13817	21820-21824	FW	denotes	vivo
T13815	21825-21830	NNS	denotes	roles
T13818	21830-21831	.	denotes	.
T13819	21831-21933	sentence	denotes	The two enzymes share 62% sequence identity and are very similar in their structure and biochemistry.
T13820	21832-21835	DT	denotes	The
T13822	21836-21839	CD	denotes	two
T13821	21840-21847	NNS	denotes	enzymes
T13823	21848-21853	VBP	denotes	share
T13824	21854-21856	CD	denotes	62
T13825	21856-21857	NN	denotes	%
T13827	21858-21866	NN	denotes	sequence
T13826	21867-21875	NN	denotes	identity
T13828	21876-21879	CC	denotes	and
T13829	21880-21883	VBP	denotes	are
T13830	21884-21888	RB	denotes	very
T13831	21889-21896	JJ	denotes	similar
T13832	21897-21899	IN	denotes	in
T13833	21900-21905	PRP$	denotes	their
T13834	21906-21915	NN	denotes	structure
T13835	21916-21919	CC	denotes	and
T13836	21920-21932	NN	denotes	biochemistry
T13837	21932-21933	.	denotes	.
T13838	21933-22084	sentence	denotes	Notably, they cleave essentially the same set of substrates in vitro, suggesting that they have the potential to carry out the same functions in vivo.
T13839	21934-21941	RB	denotes	Notably
T13841	21941-21943	,	denotes	,
T13842	21943-21947	PRP	denotes	they
T13840	21948-21954	VBP	denotes	cleave
T13843	21955-21966	RB	denotes	essentially
T13844	21967-21970	DT	denotes	the
T13846	21971-21975	JJ	denotes	same
T13845	21976-21979	NN	denotes	set
T13847	21980-21982	IN	denotes	of
T13848	21983-21993	NNS	denotes	substrates
T13849	21994-21996	FW	denotes	in
T13850	21997-22002	FW	denotes	vitro
T13851	22002-22004	,	denotes	,
T13852	22004-22014	VBG	denotes	suggesting
T13853	22015-22019	IN	denotes	that
T13855	22020-22024	PRP	denotes	they
T13854	22025-22029	VBP	denotes	have
T13856	22030-22033	DT	denotes	the
T13857	22034-22043	NN	denotes	potential
T13858	22044-22046	TO	denotes	to
T13859	22047-22052	VB	denotes	carry
T13860	22053-22056	RP	denotes	out
T13861	22057-22060	DT	denotes	the
T13863	22061-22065	JJ	denotes	same
T13862	22066-22075	NNS	denotes	functions
T13864	22076-22078	FW	denotes	in
T13865	22079-22083	FW	denotes	vivo
T13866	22083-22084	.	denotes	.
T13867	22084-22253	sentence	denotes	On the other hand, since they require different amounts of Ca2+ for in vitro activation, it is possible that the two isoforms are differentially regulated inside cells.
T13868	22085-22087	IN	denotes	On
T13870	22088-22091	DT	denotes	the
T13872	22092-22097	JJ	denotes	other
T13871	22098-22102	NN	denotes	hand
T13873	22102-22104	,	denotes	,
T13874	22104-22109	IN	denotes	since
T13876	22110-22114	PRP	denotes	they
T13875	22115-22122	VBP	denotes	require
T13877	22123-22132	JJ	denotes	different
T13878	22133-22140	NNS	denotes	amounts
T13879	22141-22143	IN	denotes	of
T13880	22144-22147	NN	denotes	Ca2
T13881	22147-22148	SYM	denotes	+
T13882	22149-22152	IN	denotes	for
T13883	22153-22155	FW	denotes	in
T13884	22156-22161	FW	denotes	vitro
T13885	22162-22172	NN	denotes	activation
T13886	22172-22174	,	denotes	,
T13887	22174-22176	PRP	denotes	it
T13869	22177-22179	VBZ	denotes	is
T13888	22180-22188	JJ	denotes	possible
T13889	22189-22193	IN	denotes	that
T13891	22194-22197	DT	denotes	the
T13893	22198-22201	CD	denotes	two
T13892	22202-22210	NNS	denotes	isoforms
T13894	22211-22214	VBP	denotes	are
T13895	22215-22229	RB	denotes	differentially
T13890	22230-22239	VBN	denotes	regulated
T13896	22240-22246	IN	denotes	inside
T13897	22247-22252	NNS	denotes	cells
T13898	22252-22253	.	denotes	.
T13899	22253-22406	sentence	denotes	It is now clear, from the gene targeting work done in mice, that μ- and m-calpain have some distinct physiological roles, at least during embryogenesis.
T13900	22254-22256	PRP	denotes	It
T13901	22257-22259	VBZ	denotes	is
T13902	22260-22263	RB	denotes	now
T13903	22264-22269	JJ	denotes	clear
T13904	22269-22271	,	denotes	,
T13905	22271-22275	IN	denotes	from
T13906	22276-22279	DT	denotes	the
T13908	22280-22284	NN	denotes	gene
T13909	22285-22294	NN	denotes	targeting
T13907	22295-22299	NN	denotes	work
T13910	22300-22304	VBN	denotes	done
T13911	22305-22307	IN	denotes	in
T13912	22308-22312	NNS	denotes	mice
T13913	22312-22314	,	denotes	,
T13914	22314-22318	IN	denotes	that
T13916	22319-22320	NN	denotes	μ
T13918	22320-22321	HYPH	denotes	-
T13919	22322-22325	CC	denotes	and
T13920	22326-22327	NN	denotes	m
T13921	22327-22328	HYPH	denotes	-
T13917	22328-22335	NN	denotes	calpain
T13915	22336-22340	VBP	denotes	have
T13922	22341-22345	DT	denotes	some
T13924	22346-22354	JJ	denotes	distinct
T13925	22355-22368	JJ	denotes	physiological
T13923	22369-22374	NNS	denotes	roles
T13926	22374-22376	,	denotes	,
T13927	22376-22378	RB	denotes	at
T13928	22379-22384	RBS	denotes	least
T13929	22385-22391	IN	denotes	during
T13930	22392-22405	NN	denotes	embryogenesis
T13931	22405-22406	.	denotes	.
T13932	22406-22586	sentence	denotes	As noted, whereas Capn2 null murine embryos die prior to implantation, homozygous disruption of the μ-calpain large subunit gene, Capn1, did not affect the viability of mice [33].
T13933	22407-22409	IN	denotes	As
T13934	22410-22415	VBN	denotes	noted
T13936	22415-22417	,	denotes	,
T13937	22417-22424	IN	denotes	whereas
T13939	22425-22430	NN	denotes	Capn2
T13940	22431-22435	JJ	denotes	null
T13942	22436-22442	JJ	denotes	murine
T13941	22443-22450	NNS	denotes	embryos
T13938	22451-22454	VBP	denotes	die
T13943	22455-22460	JJ	denotes	prior
T13944	22461-22463	IN	denotes	to
T13945	22464-22476	NN	denotes	implantation
T13946	22476-22478	,	denotes	,
T13947	22478-22488	JJ	denotes	homozygous
T13948	22489-22499	NN	denotes	disruption
T13949	22500-22502	IN	denotes	of
T13950	22503-22506	DT	denotes	the
T13952	22507-22508	NN	denotes	μ
T13954	22508-22509	HYPH	denotes	-
T13953	22509-22516	NN	denotes	calpain
T13955	22517-22522	JJ	denotes	large
T13956	22523-22530	NN	denotes	subunit
T13951	22531-22535	NN	denotes	gene
T13957	22535-22537	,	denotes	,
T13958	22537-22542	NN	denotes	Capn1
T13959	22542-22544	,	denotes	,
T13960	22544-22547	VBD	denotes	did
T13961	22548-22551	RB	denotes	not
T13935	22552-22558	VB	denotes	affect
T13962	22559-22562	DT	denotes	the
T13963	22563-22572	NN	denotes	viability
T13964	22573-22575	IN	denotes	of
T13965	22576-22580	NNS	denotes	mice
T13966	22581-22582	-LRB-	denotes	[
T13967	22582-22584	CD	denotes	33
T13968	22584-22585	-RRB-	denotes	]
T13969	22585-22586	.	denotes	.
T13970	22586-22803	sentence	denotes	The principal phenotype observed as a result of Capn1 deficiency involved a disturbance in platelet function, possibly at the level of the tyrosine phosphorylation of certain proteins involved in platelet activation.
T13971	22587-22590	DT	denotes	The
T13973	22591-22600	JJ	denotes	principal
T13972	22601-22610	NN	denotes	phenotype
T13975	22611-22619	VBN	denotes	observed
T13976	22620-22622	IN	denotes	as
T13977	22623-22624	DT	denotes	a
T13978	22625-22631	NN	denotes	result
T13979	22632-22634	IN	denotes	of
T13980	22635-22640	NN	denotes	Capn1
T13981	22641-22651	NN	denotes	deficiency
T13974	22652-22660	VBD	denotes	involved
T13982	22661-22662	DT	denotes	a
T13983	22663-22674	NN	denotes	disturbance
T13984	22675-22677	IN	denotes	in
T13985	22678-22686	NN	denotes	platelet
T13986	22687-22695	NN	denotes	function
T13987	22695-22697	,	denotes	,
T13988	22697-22705	RB	denotes	possibly
T13989	22706-22708	IN	denotes	at
T13990	22709-22712	DT	denotes	the
T13991	22713-22718	NN	denotes	level
T13992	22719-22721	IN	denotes	of
T13993	22722-22725	DT	denotes	the
T13995	22726-22734	NN	denotes	tyrosine
T13994	22735-22750	NN	denotes	phosphorylation
T13996	22751-22753	IN	denotes	of
T13997	22754-22761	JJ	denotes	certain
T13998	22762-22770	NN	denotes	proteins
T13999	22771-22779	VBN	denotes	involved
T14000	22780-22782	IN	denotes	in
T14001	22783-22791	NN	denotes	platelet
T14002	22792-22802	NN	denotes	activation
T14003	22802-22803	.	denotes	.
T14004	22803-23013	sentence	denotes	Both μ- and m-calpain activities are present in most mammalian cells, although the published data, owing to weaknesses in the available methodology, do not provide reliable estimates of their relative amounts.
T14005	22804-22808	CC	denotes	Both
T14007	22809-22810	NN	denotes	μ
T14009	22810-22811	HYPH	denotes	-
T14010	22812-22815	CC	denotes	and
T14011	22816-22817	NN	denotes	m
T14012	22817-22818	HYPH	denotes	-
T14008	22818-22825	NN	denotes	calpain
T14006	22826-22836	NNS	denotes	activities
T14013	22837-22840	VBP	denotes	are
T14014	22841-22848	JJ	denotes	present
T14015	22849-22851	IN	denotes	in
T14016	22852-22856	JJS	denotes	most
T14018	22857-22866	JJ	denotes	mammalian
T14017	22867-22872	NNS	denotes	cells
T14019	22872-22874	,	denotes	,
T14020	22874-22882	IN	denotes	although
T14022	22883-22886	DT	denotes	the
T14024	22887-22896	VBN	denotes	published
T14023	22897-22901	NNS	denotes	data
T14025	22901-22903	,	denotes	,
T14026	22903-22908	VBG	denotes	owing
T14027	22909-22911	IN	denotes	to
T14028	22912-22922	NNS	denotes	weaknesses
T14029	22923-22925	IN	denotes	in
T14030	22926-22929	DT	denotes	the
T14032	22930-22939	JJ	denotes	available
T14031	22940-22951	NN	denotes	methodology
T14033	22951-22953	,	denotes	,
T14034	22953-22955	VBP	denotes	do
T14035	22956-22959	RB	denotes	not
T14021	22960-22967	VB	denotes	provide
T14036	22968-22976	JJ	denotes	reliable
T14037	22977-22986	NNS	denotes	estimates
T14038	22987-22989	IN	denotes	of
T14039	22990-22995	PRP$	denotes	their
T14041	22996-23004	JJ	denotes	relative
T14040	23005-23012	NNS	denotes	amounts
T14042	23012-23013	.	denotes	.
T14043	23013-23124	sentence	denotes	Platelets and erythrocytes contain abundant μ-calpain activity, while m-calpain activity is barely detectable.
T14044	23014-23023	NNS	denotes	Platelets
T14046	23024-23027	CC	denotes	and
T14047	23028-23040	NNS	denotes	erythrocytes
T14045	23041-23048	VBP	denotes	contain
T14048	23049-23057	JJ	denotes	abundant
T14050	23058-23059	NN	denotes	μ
T14052	23059-23060	HYPH	denotes	-
T14051	23060-23067	NN	denotes	calpain
T14049	23068-23076	NN	denotes	activity
T14053	23076-23078	,	denotes	,
T14054	23078-23083	IN	denotes	while
T14056	23084-23085	NN	denotes	m
T14058	23085-23086	HYPH	denotes	-
T14057	23086-23093	NN	denotes	calpain
T14059	23094-23102	NN	denotes	activity
T14055	23103-23105	VBZ	denotes	is
T14060	23106-23112	RB	denotes	barely
T14061	23113-23123	JJ	denotes	detectable
T14062	23123-23124	.	denotes	.
T14063	23124-23256	sentence	denotes	Compensation for μ-calpain deficiency by m-calpain is therefore less likely in platelets and erythrocytes than in other cell types.
T14064	23125-23137	NN	denotes	Compensation
T14066	23138-23141	IN	denotes	for
T14067	23142-23143	NN	denotes	μ
T14069	23143-23144	HYPH	denotes	-
T14068	23144-23151	NN	denotes	calpain
T14070	23152-23162	NN	denotes	deficiency
T14071	23163-23165	IN	denotes	by
T14072	23166-23167	NN	denotes	m
T14074	23167-23168	HYPH	denotes	-
T14073	23168-23175	NN	denotes	calpain
T14065	23176-23178	VBZ	denotes	is
T14075	23179-23188	RB	denotes	therefore
T14076	23189-23193	RBR	denotes	less
T14077	23194-23200	JJ	denotes	likely
T14078	23201-23203	IN	denotes	in
T14079	23204-23213	NNS	denotes	platelets
T14080	23214-23217	CC	denotes	and
T14081	23218-23230	NNS	denotes	erythrocytes
T14082	23231-23235	IN	denotes	than
T14083	23236-23238	IN	denotes	in
T14084	23239-23244	JJ	denotes	other
T14086	23245-23249	NN	denotes	cell
T14085	23250-23255	NNS	denotes	types
T14087	23255-23256	.	denotes	.
T14088	23256-23494	sentence	denotes	As a result, it is not possible to determine whether the absence of marked phenotype in Capn1-/- animals is due to a compensatory affect by the remaining m-calpain activity, or whether the functions of μ-calpain are simply not essential.
T14089	23257-23259	IN	denotes	As
T14091	23260-23261	DT	denotes	a
T14092	23262-23268	NN	denotes	result
T14093	23268-23270	,	denotes	,
T14094	23270-23272	PRP	denotes	it
T14090	23273-23275	VBZ	denotes	is
T14095	23276-23279	RB	denotes	not
T14096	23280-23288	JJ	denotes	possible
T14097	23289-23291	TO	denotes	to
T14098	23292-23301	VB	denotes	determine
T14099	23302-23309	IN	denotes	whether
T14101	23310-23313	DT	denotes	the
T14102	23314-23321	NN	denotes	absence
T14103	23322-23324	IN	denotes	of
T14104	23325-23331	JJ	denotes	marked
T14105	23332-23341	NN	denotes	phenotype
T14106	23342-23344	IN	denotes	in
T14107	23345-23350	NN	denotes	Capn1
T14109	23350-23351	SYM	denotes	-
T14110	23351-23352	HYPH	denotes	/
T14111	23352-23353	SYM	denotes	-
T14108	23354-23361	NNS	denotes	animals
T14100	23362-23364	VBZ	denotes	is
T14112	23365-23368	IN	denotes	due
T14113	23369-23371	IN	denotes	to
T14114	23372-23373	DT	denotes	a
T14116	23374-23386	JJ	denotes	compensatory
T14115	23387-23393	NN	denotes	affect
T14117	23394-23396	IN	denotes	by
T14118	23397-23400	DT	denotes	the
T14120	23401-23410	VBG	denotes	remaining
T14121	23411-23412	NN	denotes	m
T14123	23412-23413	HYPH	denotes	-
T14122	23413-23420	NN	denotes	calpain
T14119	23421-23429	NN	denotes	activity
T14124	23429-23431	,	denotes	,
T14125	23431-23433	CC	denotes	or
T14126	23434-23441	IN	denotes	whether
T14128	23442-23445	DT	denotes	the
T14129	23446-23455	NNS	denotes	functions
T14130	23456-23458	IN	denotes	of
T14131	23459-23460	NN	denotes	μ
T14133	23460-23461	HYPH	denotes	-
T14132	23461-23468	NN	denotes	calpain
T14127	23469-23472	VBP	denotes	are
T14134	23473-23479	RB	denotes	simply
T14135	23480-23483	RB	denotes	not
T14136	23484-23493	JJ	denotes	essential
T14137	23493-23494	.	denotes	.
T14138	23494-23638	sentence	denotes	In contrast, lethality in Capn2-/- embryos demonstrates that m-calpain activity is essential for embryonic development beyond the 8-cell stage.
T14139	23495-23497	IN	denotes	In
T14141	23498-23506	NN	denotes	contrast
T14142	23506-23508	,	denotes	,
T14143	23508-23517	NN	denotes	lethality
T14144	23518-23520	IN	denotes	in
T14145	23521-23526	NN	denotes	Capn2
T14147	23526-23527	SYM	denotes	-
T14148	23527-23528	HYPH	denotes	/
T14149	23528-23529	SYM	denotes	-
T14146	23530-23537	NNS	denotes	embryos
T14140	23538-23550	VBZ	denotes	demonstrates
T14150	23551-23555	IN	denotes	that
T14152	23556-23557	NN	denotes	m
T14154	23557-23558	HYPH	denotes	-
T14153	23558-23565	NN	denotes	calpain
T14155	23566-23574	NN	denotes	activity
T14151	23575-23577	VBZ	denotes	is
T14156	23578-23587	JJ	denotes	essential
T14157	23588-23591	IN	denotes	for
T14158	23592-23601	JJ	denotes	embryonic
T14159	23602-23613	NN	denotes	development
T14160	23614-23620	IN	denotes	beyond
T14161	23621-23624	DT	denotes	the
T14163	23625-23626	CD	denotes	8
T14165	23626-23627	HYPH	denotes	-
T14164	23627-23631	NN	denotes	cell
T14162	23632-23637	NN	denotes	stage
T14166	23637-23638	.	denotes	.
T14167	23638-23719	sentence	denotes	It follows that at least some functions of μ-calpain and m-calpain are distinct.
T14168	23639-23641	PRP	denotes	It
T14169	23642-23649	VBZ	denotes	follows
T14170	23650-23654	IN	denotes	that
T14172	23655-23657	RB	denotes	at
T14173	23658-23663	RBS	denotes	least
T14175	23664-23668	DT	denotes	some
T14174	23669-23678	NNS	denotes	functions
T14176	23679-23681	IN	denotes	of
T14177	23682-23683	NN	denotes	μ
T14179	23683-23684	HYPH	denotes	-
T14178	23684-23691	NN	denotes	calpain
T14180	23692-23695	CC	denotes	and
T14181	23696-23697	NN	denotes	m
T14183	23697-23698	HYPH	denotes	-
T14182	23698-23705	NN	denotes	calpain
T14171	23706-23709	VBP	denotes	are
T14184	23710-23718	JJ	denotes	distinct
T14185	23718-23719	.	denotes	.
T14186	23719-23821	sentence	denotes	The underlying cause of the preimplantation lethality in Capn2-/- embryos has not yet been clarified.
T14187	23720-23723	DT	denotes	The
T14189	23724-23734	VBG	denotes	underlying
T14188	23735-23740	NN	denotes	cause
T14191	23741-23743	IN	denotes	of
T14192	23744-23747	DT	denotes	the
T14194	23748-23763	NN	denotes	preimplantation
T14193	23764-23773	NN	denotes	lethality
T14195	23774-23776	IN	denotes	in
T14196	23777-23782	NN	denotes	Capn2
T14198	23782-23783	SYM	denotes	-
T14199	23783-23784	HYPH	denotes	/
T14200	23784-23785	SYM	denotes	-
T14197	23786-23793	NNS	denotes	embryos
T14201	23794-23797	VBZ	denotes	has
T14202	23798-23801	RB	denotes	not
T14203	23802-23805	RB	denotes	yet
T14204	23806-23810	VBN	denotes	been
T14190	23811-23820	VBN	denotes	clarified
T14205	23820-23821	.	denotes	.
T14206	23821-23935	sentence	denotes	The two homozygous null embryos identified at the 8-cell stage did not present any obvious morphological defects.
T14207	23822-23825	DT	denotes	The
T14209	23826-23829	CD	denotes	two
T14210	23830-23840	JJ	denotes	homozygous
T14211	23841-23845	JJ	denotes	null
T14208	23846-23853	NNS	denotes	embryos
T14213	23854-23864	VBN	denotes	identified
T14214	23865-23867	IN	denotes	at
T14215	23868-23871	DT	denotes	the
T14217	23872-23873	CD	denotes	8
T14219	23873-23874	HYPH	denotes	-
T14218	23874-23878	NN	denotes	cell
T14216	23879-23884	NN	denotes	stage
T14220	23885-23888	VBD	denotes	did
T14221	23889-23892	RB	denotes	not
T14212	23893-23900	VB	denotes	present
T14222	23901-23904	DT	denotes	any
T14224	23905-23912	JJ	denotes	obvious
T14225	23913-23926	JJ	denotes	morphological
T14223	23927-23934	NNS	denotes	defects
T14226	23934-23935	.	denotes	.
T14227	23935-24071	sentence	denotes	However, the fact that only two out of 90 successfully genotyped pre-implantation embryos proved to be Capn2-/- is in itself revealing.
T14228	23936-23943	RB	denotes	However
T14230	23943-23945	,	denotes	,
T14231	23945-23948	DT	denotes	the
T14232	23949-23953	NN	denotes	fact
T14233	23954-23958	IN	denotes	that
T14235	23959-23963	RB	denotes	only
T14237	23964-23967	CD	denotes	two
T14239	23968-23971	IN	denotes	out
T14240	23972-23974	IN	denotes	of
T14238	23975-23977	CD	denotes	90
T14241	23978-23990	RB	denotes	successfully
T14242	23991-24000	VBN	denotes	genotyped
T14243	24001-24017	JJ	denotes	pre-implantation
T14236	24018-24025	NNS	denotes	embryos
T14234	24026-24032	VBD	denotes	proved
T14244	24033-24035	TO	denotes	to
T14245	24036-24038	VB	denotes	be
T14246	24039-24044	NN	denotes	Capn2
T14247	24044-24045	SYM	denotes	-
T14248	24045-24046	HYPH	denotes	/
T14249	24046-24047	SYM	denotes	-
T14229	24048-24050	VBZ	denotes	is
T14250	24051-24053	IN	denotes	in
T14251	24054-24060	PRP	denotes	itself
T14252	24061-24070	VBG	denotes	revealing
T14253	24070-24071	.	denotes	.
T14254	24071-24178	sentence	denotes	Preimplantation lethality resulting from gene knockouts can typically be attributed to two general causes.
T14255	24072-24087	NN	denotes	Preimplantation
T14256	24088-24097	NN	denotes	lethality
T14258	24098-24107	VBG	denotes	resulting
T14259	24108-24112	IN	denotes	from
T14260	24113-24117	NN	denotes	gene
T14261	24118-24127	NNS	denotes	knockouts
T14262	24128-24131	MD	denotes	can
T14263	24132-24141	RB	denotes	typically
T14264	24142-24144	VB	denotes	be
T14257	24145-24155	VBN	denotes	attributed
T14265	24156-24158	IN	denotes	to
T14266	24159-24162	CD	denotes	two
T14268	24163-24170	JJ	denotes	general
T14267	24171-24177	NNS	denotes	causes
T14269	24177-24178	.	denotes	.
T14270	24178-24305	sentence	denotes	In some cases, defects are incurred in the embryonic differentiation program which can often be observed morphologically [38].
T14271	24179-24181	IN	denotes	In
T14273	24182-24186	DT	denotes	some
T14274	24187-24192	NNS	denotes	cases
T14275	24192-24194	,	denotes	,
T14276	24194-24201	NNS	denotes	defects
T14277	24202-24205	VBP	denotes	are
T14272	24206-24214	VBN	denotes	incurred
T14278	24215-24217	IN	denotes	in
T14279	24218-24221	DT	denotes	the
T14281	24222-24231	JJ	denotes	embryonic
T14282	24232-24247	NN	denotes	differentiation
T14280	24248-24255	NN	denotes	program
T14283	24256-24261	WDT	denotes	which
T14285	24262-24265	MD	denotes	can
T14286	24266-24271	RB	denotes	often
T14287	24272-24274	VB	denotes	be
T14284	24275-24283	VBN	denotes	observed
T14288	24284-24299	RB	denotes	morphologically
T14289	24300-24301	-LRB-	denotes	[
T14290	24301-24303	CD	denotes	38
T14291	24303-24304	-RRB-	denotes	]
T14292	24304-24305	.	denotes	.
T14293	24305-24440	sentence	denotes	Null embryos of this type often survive beyond the morula stage, and a Mendelian distribution of embryonic genotypes is usually noted.
T14294	24306-24310	JJ	denotes	Null
T14295	24311-24318	NNS	denotes	embryos
T14297	24319-24321	IN	denotes	of
T14298	24322-24326	DT	denotes	this
T14299	24327-24331	NN	denotes	type
T14300	24332-24337	RB	denotes	often
T14296	24338-24345	VBP	denotes	survive
T14301	24346-24352	IN	denotes	beyond
T14302	24353-24356	DT	denotes	the
T14304	24357-24363	NN	denotes	morula
T14303	24364-24369	NN	denotes	stage
T14305	24369-24371	,	denotes	,
T14306	24371-24374	CC	denotes	and
T14307	24375-24376	DT	denotes	a
T14309	24377-24386	JJ	denotes	Mendelian
T14308	24387-24399	NN	denotes	distribution
T14311	24400-24402	IN	denotes	of
T14312	24403-24412	JJ	denotes	embryonic
T14313	24413-24422	NNS	denotes	genotypes
T14314	24423-24425	VBZ	denotes	is
T14315	24426-24433	RB	denotes	usually
T14310	24434-24439	VBN	denotes	noted
T14316	24439-24440	.	denotes	.
T14317	24440-24572	sentence	denotes	In other cases, however, the gene disruption is thought to compromise fatally the viability of cells in the early embryo [3,39-43].
T14318	24441-24443	IN	denotes	In
T14320	24444-24449	JJ	denotes	other
T14321	24450-24455	NNS	denotes	cases
T14322	24455-24457	,	denotes	,
T14323	24457-24464	RB	denotes	however
T14324	24464-24466	,	denotes	,
T14325	24466-24469	DT	denotes	the
T14327	24470-24474	NN	denotes	gene
T14326	24475-24485	NN	denotes	disruption
T14328	24486-24488	VBZ	denotes	is
T14319	24489-24496	VBN	denotes	thought
T14329	24497-24499	TO	denotes	to
T14330	24500-24510	VB	denotes	compromise
T14331	24511-24518	RB	denotes	fatally
T14332	24519-24522	DT	denotes	the
T14333	24523-24532	NN	denotes	viability
T14334	24533-24535	IN	denotes	of
T14335	24536-24541	NNS	denotes	cells
T14336	24542-24544	IN	denotes	in
T14337	24545-24548	DT	denotes	the
T14339	24549-24554	JJ	denotes	early
T14338	24555-24561	NN	denotes	embryo
T14340	24562-24563	-LRB-	denotes	[
T14342	24563-24564	CD	denotes	3
T14343	24564-24565	,	denotes	,
T14341	24565-24567	CD	denotes	39
T14344	24567-24568	SYM	denotes	-
T14345	24568-24570	CD	denotes	43
T14346	24570-24571	-RRB-	denotes	]
T14347	24571-24572	.	denotes	.
T14348	24572-24688	sentence	denotes	In these instances, only a few null embryos are ever observed and homozygous mutant ES cells could not be isolated.
T14349	24573-24575	IN	denotes	In
T14351	24576-24581	DT	denotes	these
T14352	24582-24591	NNS	denotes	instances
T14353	24591-24593	,	denotes	,
T14354	24593-24597	RB	denotes	only
T14356	24598-24599	DT	denotes	a
T14357	24600-24603	JJ	denotes	few
T14358	24604-24608	JJ	denotes	null
T14355	24609-24616	NNS	denotes	embryos
T14359	24617-24620	VBP	denotes	are
T14360	24621-24625	RB	denotes	ever
T14350	24626-24634	VBN	denotes	observed
T14361	24635-24638	CC	denotes	and
T14362	24639-24649	JJ	denotes	homozygous
T14364	24650-24656	NN	denotes	mutant
T14365	24657-24659	NN	denotes	ES
T14363	24660-24665	NNS	denotes	cells
T14367	24666-24671	MD	denotes	could
T14368	24672-24675	RB	denotes	not
T14369	24676-24678	VB	denotes	be
T14366	24679-24687	VBN	denotes	isolated
T14370	24687-24688	.	denotes	.
T14371	24688-24738	sentence	denotes	The Capn2 knockout fits into the latter category.
T14372	24689-24692	DT	denotes	The
T14374	24693-24698	NN	denotes	Capn2
T14373	24699-24707	NN	denotes	knockout
T14375	24708-24712	VBZ	denotes	fits
T14376	24713-24717	IN	denotes	into
T14377	24718-24721	DT	denotes	the
T14379	24722-24728	JJ	denotes	latter
T14378	24729-24737	NN	denotes	category
T14380	24737-24738	.	denotes	.
T14381	24738-24987	sentence	denotes	If it is true that m-calpain is essential for some aspect of cell viability, the survival of a few Capn2-/- embryos to the 8-cell stage is most likely due to the persistence of some maternal m-calpain mRNA and/or protein through 2–3 cell divisions.
T14382	24739-24741	IN	denotes	If
T14384	24742-24744	PRP	denotes	it
T14383	24745-24747	VBZ	denotes	is
T14386	24748-24752	JJ	denotes	true
T14387	24753-24757	IN	denotes	that
T14389	24758-24759	NN	denotes	m
T14391	24759-24760	HYPH	denotes	-
T14390	24760-24767	NN	denotes	calpain
T14388	24768-24770	VBZ	denotes	is
T14392	24771-24780	JJ	denotes	essential
T14393	24781-24784	IN	denotes	for
T14394	24785-24789	DT	denotes	some
T14395	24790-24796	NN	denotes	aspect
T14396	24797-24799	IN	denotes	of
T14397	24800-24804	NN	denotes	cell
T14398	24805-24814	NN	denotes	viability
T14399	24814-24816	,	denotes	,
T14400	24816-24819	DT	denotes	the
T14401	24820-24828	NN	denotes	survival
T14402	24829-24831	IN	denotes	of
T14403	24832-24833	DT	denotes	a
T14405	24834-24837	JJ	denotes	few
T14406	24838-24843	NN	denotes	Capn2
T14407	24843-24844	SYM	denotes	-
T14408	24844-24845	HYPH	denotes	/
T14409	24845-24846	SYM	denotes	-
T14404	24847-24854	NNS	denotes	embryos
T14410	24855-24857	IN	denotes	to
T14411	24858-24861	DT	denotes	the
T14413	24862-24863	CD	denotes	8
T14415	24863-24864	HYPH	denotes	-
T14414	24864-24868	NN	denotes	cell
T14412	24869-24874	NN	denotes	stage
T14385	24875-24877	VBZ	denotes	is
T14416	24878-24882	RBS	denotes	most
T14417	24883-24889	RB	denotes	likely
T14418	24890-24893	IN	denotes	due
T14419	24894-24896	IN	denotes	to
T14420	24897-24900	DT	denotes	the
T14421	24901-24912	NN	denotes	persistence
T14422	24913-24915	IN	denotes	of
T14423	24916-24920	DT	denotes	some
T14425	24921-24929	JJ	denotes	maternal
T14426	24930-24931	NN	denotes	m
T14427	24931-24932	HYPH	denotes	-
T14424	24932-24939	NN	denotes	calpain
T14428	24940-24944	NN	denotes	mRNA
T14429	24945-24948	CC	denotes	and
T14430	24948-24949	HYPH	denotes	/
T14431	24949-24951	CC	denotes	or
T14432	24952-24959	NN	denotes	protein
T14433	24960-24967	IN	denotes	through
T14434	24968-24969	CD	denotes	2
T14436	24969-24970	SYM	denotes	–
T14435	24970-24971	CD	denotes	3
T14438	24972-24976	NN	denotes	cell
T14437	24977-24986	NNS	denotes	divisions
T14439	24986-24987	.	denotes	.
T14440	24987-25072	sentence	denotes	The mechanistic reasons for lethality in the absence of m-calpain are still unclear.
T14441	24988-24991	DT	denotes	The
T14443	24992-25003	JJ	denotes	mechanistic
T14442	25004-25011	NNS	denotes	reasons
T14445	25012-25015	IN	denotes	for
T14446	25016-25025	NN	denotes	lethality
T14447	25026-25028	IN	denotes	in
T14448	25029-25032	DT	denotes	the
T14449	25033-25040	NN	denotes	absence
T14450	25041-25043	IN	denotes	of
T14451	25044-25045	NN	denotes	m
T14453	25045-25046	HYPH	denotes	-
T14452	25046-25053	NN	denotes	calpain
T14444	25054-25057	VBP	denotes	are
T14454	25058-25063	RB	denotes	still
T14455	25064-25071	JJ	denotes	unclear
T14456	25071-25072	.	denotes	.
T14457	25072-25292	sentence	denotes	There have been several reports of m-calpain involvement in cell proliferation in certain circumstances, including reports of its involvement in chromosome segregation during mitosis [44] as well as during meiosis [45].
T14458	25073-25078	EX	denotes	There
T14460	25079-25083	VBP	denotes	have
T14459	25084-25088	VBN	denotes	been
T14461	25089-25096	JJ	denotes	several
T14462	25097-25104	NNS	denotes	reports
T14463	25105-25107	IN	denotes	of
T14464	25108-25109	NN	denotes	m
T14466	25109-25110	HYPH	denotes	-
T14465	25110-25117	NN	denotes	calpain
T14467	25118-25129	NN	denotes	involvement
T14468	25130-25132	IN	denotes	in
T14469	25133-25137	NN	denotes	cell
T14470	25138-25151	NN	denotes	proliferation
T14471	25152-25154	IN	denotes	in
T14472	25155-25162	JJ	denotes	certain
T14473	25163-25176	NNS	denotes	circumstances
T14474	25176-25178	,	denotes	,
T14475	25178-25187	VBG	denotes	including
T14476	25188-25195	NNS	denotes	reports
T14477	25196-25198	IN	denotes	of
T14478	25199-25202	PRP$	denotes	its
T14479	25203-25214	NN	denotes	involvement
T14480	25215-25217	IN	denotes	in
T14481	25218-25228	NN	denotes	chromosome
T14482	25229-25240	NN	denotes	segregation
T14483	25241-25247	IN	denotes	during
T14484	25248-25255	NN	denotes	mitosis
T14485	25256-25257	-LRB-	denotes	[
T14486	25257-25259	CD	denotes	44
T14487	25259-25260	-RRB-	denotes	]
T14488	25261-25263	RB	denotes	as
T14490	25264-25268	RB	denotes	well
T14489	25269-25271	IN	denotes	as
T14491	25272-25278	IN	denotes	during
T14492	25279-25286	NN	denotes	meiosis
T14493	25287-25288	-LRB-	denotes	[
T14494	25288-25290	CD	denotes	45
T14495	25290-25291	-RRB-	denotes	]
T14496	25291-25292	.	denotes	.
T14497	25292-25396	sentence	denotes	Defects in migration, reported in Capn4-/- cells [9], could contribute to failed embryonic development.
T14498	25293-25300	NNS	denotes	Defects
T14500	25301-25303	IN	denotes	in
T14501	25304-25313	NN	denotes	migration
T14502	25313-25315	,	denotes	,
T14503	25315-25323	VBN	denotes	reported
T14504	25324-25326	IN	denotes	in
T14505	25327-25332	NN	denotes	Capn4
T14507	25332-25333	SYM	denotes	-
T14508	25333-25334	HYPH	denotes	/
T14509	25334-25335	SYM	denotes	-
T14506	25336-25341	NNS	denotes	cells
T14510	25342-25343	-LRB-	denotes	[
T14511	25343-25344	CD	denotes	9
T14512	25344-25345	-RRB-	denotes	]
T14513	25345-25347	,	denotes	,
T14514	25347-25352	MD	denotes	could
T14499	25353-25363	VB	denotes	contribute
T14515	25364-25366	IN	denotes	to
T14516	25367-25373	VBN	denotes	failed
T14518	25374-25383	JJ	denotes	embryonic
T14517	25384-25395	NN	denotes	development
T14519	25395-25396	.	denotes	.
T14520	25396-25645	sentence	denotes	The association between calpain and cell viability has been noted in Capn4-/- MEFs and other cell lines, although the reported work frequently did not distinguish between μ- and m-calpain, and did not show that calpain was strictly essential [5,6].
T14521	25397-25400	DT	denotes	The
T14522	25401-25412	NN	denotes	association
T14524	25413-25420	IN	denotes	between
T14525	25421-25428	NN	denotes	calpain
T14526	25429-25432	CC	denotes	and
T14527	25433-25437	NN	denotes	cell
T14528	25438-25447	NN	denotes	viability
T14529	25448-25451	VBZ	denotes	has
T14530	25452-25456	VBN	denotes	been
T14523	25457-25462	VBN	denotes	noted
T14531	25463-25465	IN	denotes	in
T14532	25466-25471	NN	denotes	Capn4
T14534	25471-25472	SYM	denotes	-
T14535	25472-25473	HYPH	denotes	/
T14536	25473-25474	SYM	denotes	-
T14533	25475-25479	NNS	denotes	MEFs
T14537	25480-25483	CC	denotes	and
T14538	25484-25489	JJ	denotes	other
T14540	25490-25494	NN	denotes	cell
T14539	25495-25500	NNS	denotes	lines
T14541	25500-25502	,	denotes	,
T14542	25502-25510	IN	denotes	although
T14544	25511-25514	DT	denotes	the
T14546	25515-25523	VBN	denotes	reported
T14545	25524-25528	NN	denotes	work
T14547	25529-25539	RB	denotes	frequently
T14548	25540-25543	VBD	denotes	did
T14549	25544-25547	RB	denotes	not
T14543	25548-25559	VB	denotes	distinguish
T14550	25560-25567	IN	denotes	between
T14551	25568-25569	NN	denotes	μ
T14553	25569-25570	HYPH	denotes	-
T14554	25571-25574	CC	denotes	and
T14555	25575-25576	NN	denotes	m
T14556	25576-25577	HYPH	denotes	-
T14552	25577-25584	NN	denotes	calpain
T14557	25584-25586	,	denotes	,
T14558	25586-25589	CC	denotes	and
T14559	25590-25593	VBD	denotes	did
T14561	25594-25597	RB	denotes	not
T14560	25598-25602	VB	denotes	show
T14562	25603-25607	IN	denotes	that
T14564	25608-25615	NN	denotes	calpain
T14563	25616-25619	VBD	denotes	was
T14565	25620-25628	RB	denotes	strictly
T14566	25629-25638	JJ	denotes	essential
T14567	25639-25640	-LRB-	denotes	[
T14569	25640-25641	CD	denotes	5
T14570	25641-25642	,	denotes	,
T14568	25642-25643	CD	denotes	6
T14571	25643-25644	-RRB-	denotes	]
T14572	25644-25645	.	denotes	.
T14573	25645-25774	sentence	denotes	In both CHO cells and Capn4-/- MEFs, calpain was shown to influence cell proliferation, but only at very low cell densities [5].
T14574	25646-25648	IN	denotes	In
T14576	25649-25653	CC	denotes	both
T14578	25654-25657	NN	denotes	CHO
T14577	25658-25663	NNS	denotes	cells
T14579	25664-25667	CC	denotes	and
T14580	25668-25673	NN	denotes	Capn4
T14582	25673-25674	SYM	denotes	-
T14583	25674-25675	HYPH	denotes	/
T14584	25675-25676	SYM	denotes	-
T14581	25677-25681	NNS	denotes	MEFs
T14585	25681-25683	,	denotes	,
T14586	25683-25690	NN	denotes	calpain
T14587	25691-25694	VBD	denotes	was
T14575	25695-25700	VBN	denotes	shown
T14588	25701-25703	TO	denotes	to
T14589	25704-25713	VB	denotes	influence
T14590	25714-25718	NN	denotes	cell
T14591	25719-25732	NN	denotes	proliferation
T14592	25732-25734	,	denotes	,
T14593	25734-25737	CC	denotes	but
T14595	25738-25742	RB	denotes	only
T14594	25743-25745	IN	denotes	at
T14596	25746-25750	RB	denotes	very
T14597	25751-25754	JJ	denotes	low
T14599	25755-25759	NN	denotes	cell
T14598	25760-25769	NNS	denotes	densities
T14600	25770-25771	-LRB-	denotes	[
T14601	25771-25772	CD	denotes	5
T14602	25772-25773	-RRB-	denotes	]
T14603	25773-25774	.	denotes	.
T14604	25774-25868	sentence	denotes	Calpain has also been associated with progression through the G1 stage of the cell cycle [6].
T14605	25775-25782	NN	denotes	Calpain
T14607	25783-25786	VBZ	denotes	has
T14608	25787-25791	RB	denotes	also
T14609	25792-25796	VBN	denotes	been
T14606	25797-25807	VBN	denotes	associated
T14610	25808-25812	IN	denotes	with
T14611	25813-25824	NN	denotes	progression
T14612	25825-25832	IN	denotes	through
T14613	25833-25836	DT	denotes	the
T14615	25837-25839	NN	denotes	G1
T14614	25840-25845	NN	denotes	stage
T14616	25846-25848	IN	denotes	of
T14617	25849-25852	DT	denotes	the
T14619	25853-25857	NN	denotes	cell
T14618	25858-25863	NN	denotes	cycle
T14620	25864-25865	-LRB-	denotes	[
T14621	25865-25866	CD	denotes	6
T14622	25866-25867	-RRB-	denotes	]
T14623	25867-25868	.	denotes	.
T14624	25868-26034	sentence	denotes	Furthermore, some of the proteins known to be involved in cell cycle progression, such as p53, p107, cyclin D1, and p27kip1 are reputed to be calpain substrates [3].
T14625	25869-25880	RB	denotes	Furthermore
T14627	25880-25882	,	denotes	,
T14628	25882-25886	DT	denotes	some
T14629	25887-25889	IN	denotes	of
T14630	25890-25893	DT	denotes	the
T14631	25894-25902	NN	denotes	proteins
T14632	25903-25908	VBN	denotes	known
T14633	25909-25911	TO	denotes	to
T14635	25912-25914	VB	denotes	be
T14634	25915-25923	VBN	denotes	involved
T14636	25924-25926	IN	denotes	in
T14637	25927-25931	NN	denotes	cell
T14638	25932-25937	NN	denotes	cycle
T14639	25938-25949	NN	denotes	progression
T14640	25949-25951	,	denotes	,
T14641	25951-25955	JJ	denotes	such
T14642	25956-25958	IN	denotes	as
T14643	25959-25962	NN	denotes	p53
T14644	25962-25964	,	denotes	,
T14645	25964-25968	NN	denotes	p107
T14646	25968-25970	,	denotes	,
T14647	25970-25976	NN	denotes	cyclin
T14648	25977-25979	NN	denotes	D1
T14649	25979-25981	,	denotes	,
T14650	25981-25984	CC	denotes	and
T14651	25985-25992	NN	denotes	p27kip1
T14652	25993-25996	VBP	denotes	are
T14626	25997-26004	VBN	denotes	reputed
T14653	26005-26007	TO	denotes	to
T14654	26008-26010	VB	denotes	be
T14655	26011-26018	NN	denotes	calpain
T14656	26019-26029	NNS	denotes	substrates
T14657	26030-26031	-LRB-	denotes	[
T14658	26031-26032	CD	denotes	3
T14659	26032-26033	-RRB-	denotes	]
T14660	26033-26034	.	denotes	.
T14661	26034-26144	sentence	denotes	Sperm binding to the oocyte leads to increased cytoplasmic calcium which triggers the acrosome reaction [46].
T14662	26035-26040	NN	denotes	Sperm
T14663	26041-26048	NN	denotes	binding
T14665	26049-26051	IN	denotes	to
T14666	26052-26055	DT	denotes	the
T14667	26056-26062	NN	denotes	oocyte
T14664	26063-26068	VBZ	denotes	leads
T14668	26069-26071	IN	denotes	to
T14669	26072-26081	VBN	denotes	increased
T14671	26082-26093	JJ	denotes	cytoplasmic
T14670	26094-26101	NN	denotes	calcium
T14672	26102-26107	WDT	denotes	which
T14673	26108-26116	VBZ	denotes	triggers
T14674	26117-26120	DT	denotes	the
T14676	26121-26129	NN	denotes	acrosome
T14675	26130-26138	NN	denotes	reaction
T14677	26139-26140	-LRB-	denotes	[
T14678	26140-26142	CD	denotes	46
T14679	26142-26143	-RRB-	denotes	]
T14680	26143-26144	.	denotes	.
T14681	26144-26483	sentence	denotes	Both μ- and m-calpain have recently been detected in rodent sperm [47] and oocytes [45]. m-calpain was implicated in the acrosome reaction [47] which correlated with a translocation of m-calpain to the cortical membrane in oocytes where it might participate in the release of cortical granule contents required to prevent polyspermy [45].
T14682	26145-26149	CC	denotes	Both
T14683	26150-26151	NN	denotes	μ
T14685	26151-26152	HYPH	denotes	-
T14686	26153-26156	CC	denotes	and
T14687	26157-26158	NN	denotes	m
T14688	26158-26159	HYPH	denotes	-
T14684	26159-26166	NN	denotes	calpain
T14690	26167-26171	VBP	denotes	have
T14691	26172-26180	RB	denotes	recently
T14692	26181-26185	VBN	denotes	been
T14689	26186-26194	VBN	denotes	detected
T14694	26195-26197	IN	denotes	in
T14695	26198-26204	NN	denotes	rodent
T14696	26205-26210	NN	denotes	sperm
T14697	26211-26212	-LRB-	denotes	[
T14698	26212-26214	CD	denotes	47
T14699	26214-26215	-RRB-	denotes	]
T14700	26216-26219	CC	denotes	and
T14701	26220-26227	NNS	denotes	oocytes
T14702	26228-26229	-LRB-	denotes	[
T14703	26229-26231	CD	denotes	45
T14704	26231-26232	-RRB-	denotes	]
T14705	26232-26233	.	denotes	.
T14706	26234-26235	NN	denotes	m
T14708	26235-26236	HYPH	denotes	-
T14707	26236-26243	NN	denotes	calpain
T14709	26244-26247	VBD	denotes	was
T14693	26248-26258	VBN	denotes	implicated
T14710	26259-26261	IN	denotes	in
T14711	26262-26265	DT	denotes	the
T14713	26266-26274	NN	denotes	acrosome
T14712	26275-26283	NN	denotes	reaction
T14714	26284-26285	-LRB-	denotes	[
T14715	26285-26287	CD	denotes	47
T14716	26287-26288	-RRB-	denotes	]
T14717	26289-26294	WDT	denotes	which
T14718	26295-26305	VBD	denotes	correlated
T14719	26306-26310	IN	denotes	with
T14720	26311-26312	DT	denotes	a
T14721	26313-26326	NN	denotes	translocation
T14722	26327-26329	IN	denotes	of
T14723	26330-26331	NN	denotes	m
T14725	26331-26332	HYPH	denotes	-
T14724	26332-26339	NN	denotes	calpain
T14726	26340-26342	IN	denotes	to
T14727	26343-26346	DT	denotes	the
T14729	26347-26355	JJ	denotes	cortical
T14728	26356-26364	NN	denotes	membrane
T14730	26365-26367	IN	denotes	in
T14731	26368-26375	NNS	denotes	oocytes
T14732	26376-26381	WRB	denotes	where
T14734	26382-26384	PRP	denotes	it
T14735	26385-26390	MD	denotes	might
T14733	26391-26402	VB	denotes	participate
T14736	26403-26405	IN	denotes	in
T14737	26406-26409	DT	denotes	the
T14738	26410-26417	NN	denotes	release
T14739	26418-26420	IN	denotes	of
T14740	26421-26429	JJ	denotes	cortical
T14742	26430-26437	NN	denotes	granule
T14741	26438-26446	NNS	denotes	contents
T14743	26447-26455	VBN	denotes	required
T14744	26456-26458	TO	denotes	to
T14745	26459-26466	VB	denotes	prevent
T14746	26467-26477	NN	denotes	polyspermy
T14747	26478-26479	-LRB-	denotes	[
T14748	26479-26481	CD	denotes	45
T14749	26481-26482	-RRB-	denotes	]
T14750	26482-26483	.	denotes	.
T14751	26483-26619	sentence	denotes	m-calpain also relocalized to the oocyte meiotic spindle after fertilization, were it could be involved in chromosome segregation [45].
T14752	26484-26485	NN	denotes	m
T14754	26485-26486	HYPH	denotes	-
T14753	26486-26493	NN	denotes	calpain
T14756	26494-26498	RB	denotes	also
T14755	26499-26510	VBD	denotes	relocalized
T14757	26511-26513	IN	denotes	to
T14758	26514-26517	DT	denotes	the
T14760	26518-26524	NN	denotes	oocyte
T14761	26525-26532	JJ	denotes	meiotic
T14759	26533-26540	NN	denotes	spindle
T14762	26541-26546	IN	denotes	after
T14763	26547-26560	NN	denotes	fertilization
T14764	26560-26562	,	denotes	,
T14765	26562-26566	VBD	denotes	were
T14767	26567-26569	PRP	denotes	it
T14768	26570-26575	MD	denotes	could
T14769	26576-26578	VB	denotes	be
T14766	26579-26587	VBN	denotes	involved
T14770	26588-26590	IN	denotes	in
T14771	26591-26601	NN	denotes	chromosome
T14772	26602-26613	NN	denotes	segregation
T14773	26614-26615	-LRB-	denotes	[
T14774	26615-26617	CD	denotes	45
T14775	26617-26618	-RRB-	denotes	]
T14776	26618-26619	.	denotes	.
T14777	26619-26729	sentence	denotes	Polyspermy or defective chromosome segregation would both have lethal effects on early embryonic development.
T14778	26620-26630	NN	denotes	Polyspermy
T14780	26631-26633	CC	denotes	or
T14781	26634-26643	JJ	denotes	defective
T14783	26644-26654	NN	denotes	chromosome
T14782	26655-26666	NN	denotes	segregation
T14784	26667-26672	MD	denotes	would
T14785	26673-26677	RB	denotes	both
T14779	26678-26682	VB	denotes	have
T14786	26683-26689	JJ	denotes	lethal
T14787	26690-26697	NNS	denotes	effects
T14788	26698-26700	IN	denotes	on
T14789	26701-26706	JJ	denotes	early
T14791	26707-26716	JJ	denotes	embryonic
T14790	26717-26728	NN	denotes	development
T14792	26728-26729	.	denotes	.
T14793	26729-27003	sentence	denotes	It should also be stressed that Capn4-/- ES and MEF cells from the presumptive hypomorphic allele can be maintained in culture despite an apparent lack of calpain activity, as assessed by casein zymography or by the appearance of characteristic spectrin breakdown products.
T14794	26730-26732	PRP	denotes	It
T14796	26733-26739	MD	denotes	should
T14797	26740-26744	RB	denotes	also
T14798	26745-26747	VB	denotes	be
T14795	26748-26756	VBN	denotes	stressed
T14799	26757-26761	IN	denotes	that
T14801	26762-26767	NN	denotes	Capn4
T14803	26767-26768	SYM	denotes	-
T14804	26768-26769	HYPH	denotes	/
T14805	26769-26770	SYM	denotes	-
T14806	26771-26773	NN	denotes	ES
T14807	26774-26777	CC	denotes	and
T14808	26778-26781	NN	denotes	MEF
T14802	26782-26787	NNS	denotes	cells
T14809	26788-26792	IN	denotes	from
T14810	26793-26796	DT	denotes	the
T14812	26797-26808	JJ	denotes	presumptive
T14813	26809-26820	JJ	denotes	hypomorphic
T14811	26821-26827	NN	denotes	allele
T14814	26828-26831	MD	denotes	can
T14815	26832-26834	VB	denotes	be
T14800	26835-26845	VBN	denotes	maintained
T14816	26846-26848	IN	denotes	in
T14817	26849-26856	NN	denotes	culture
T14818	26857-26864	IN	denotes	despite
T14819	26865-26867	DT	denotes	an
T14821	26868-26876	JJ	denotes	apparent
T14820	26877-26881	NN	denotes	lack
T14822	26882-26884	IN	denotes	of
T14823	26885-26892	NN	denotes	calpain
T14824	26893-26901	NN	denotes	activity
T14825	26901-26903	,	denotes	,
T14826	26903-26905	IN	denotes	as
T14827	26906-26914	VBN	denotes	assessed
T14828	26915-26917	IN	denotes	by
T14829	26918-26924	NN	denotes	casein
T14830	26925-26935	NN	denotes	zymography
T14831	26936-26938	CC	denotes	or
T14832	26939-26941	IN	denotes	by
T14833	26942-26945	DT	denotes	the
T14834	26946-26956	NN	denotes	appearance
T14835	26957-26959	IN	denotes	of
T14836	26960-26974	JJ	denotes	characteristic
T14838	26975-26983	NN	denotes	spectrin
T14839	26984-26993	NN	denotes	breakdown
T14837	26994-27002	NNS	denotes	products
T14840	27002-27003	.	denotes	.
T14841	27003-27237	sentence	denotes	It is conceivable, as discussed earlier, that a trace amount of calpain activity, beneath levels detectable by these methods, is retained from this mutant Capn4 allele, and it is sufficient for maintaining the viability of the cells.
T14842	27004-27006	PRP	denotes	It
T14843	27007-27009	VBZ	denotes	is
T14844	27010-27021	JJ	denotes	conceivable
T14845	27021-27023	,	denotes	,
T14846	27023-27025	IN	denotes	as
T14847	27026-27035	VBN	denotes	discussed
T14848	27036-27043	RBR	denotes	earlier
T14849	27043-27045	,	denotes	,
T14850	27045-27049	IN	denotes	that
T14852	27050-27051	DT	denotes	a
T14854	27052-27057	NN	denotes	trace
T14853	27058-27064	NN	denotes	amount
T14855	27065-27067	IN	denotes	of
T14856	27068-27075	NN	denotes	calpain
T14857	27076-27084	NN	denotes	activity
T14858	27084-27086	,	denotes	,
T14859	27086-27093	IN	denotes	beneath
T14860	27094-27100	NNS	denotes	levels
T14861	27101-27111	JJ	denotes	detectable
T14862	27112-27114	IN	denotes	by
T14863	27115-27120	DT	denotes	these
T14864	27121-27128	NNS	denotes	methods
T14865	27128-27130	,	denotes	,
T14866	27130-27132	VBZ	denotes	is
T14851	27133-27141	VBN	denotes	retained
T14867	27142-27146	IN	denotes	from
T14868	27147-27151	DT	denotes	this
T14870	27152-27158	NN	denotes	mutant
T14871	27159-27164	NN	denotes	Capn4
T14869	27165-27171	NN	denotes	allele
T14872	27171-27173	,	denotes	,
T14873	27173-27176	CC	denotes	and
T14874	27177-27179	PRP	denotes	it
T14875	27180-27182	VBZ	denotes	is
T14876	27183-27193	JJ	denotes	sufficient
T14877	27194-27197	IN	denotes	for
T14878	27198-27209	VBG	denotes	maintaining
T14879	27210-27213	DT	denotes	the
T14880	27214-27223	NN	denotes	viability
T14881	27224-27226	IN	denotes	of
T14882	27227-27230	DT	denotes	the
T14883	27231-27236	NNS	denotes	cells
T14884	27236-27237	.	denotes	.
T14885	27237-27385	sentence	denotes	Furthermore, calpain-independent mechanisms for protecting cell viability might exist in these cell lines that are absent in early embryonic cells.
T14886	27238-27249	RB	denotes	Furthermore
T14888	27249-27251	,	denotes	,
T14889	27251-27258	NN	denotes	calpain
T14891	27258-27259	HYPH	denotes	-
T14890	27259-27270	JJ	denotes	independent
T14892	27271-27281	NNS	denotes	mechanisms
T14893	27282-27285	IN	denotes	for
T14894	27286-27296	VBG	denotes	protecting
T14895	27297-27301	NN	denotes	cell
T14896	27302-27311	NN	denotes	viability
T14897	27312-27317	MD	denotes	might
T14887	27318-27323	VB	denotes	exist
T14898	27324-27326	IN	denotes	in
T14899	27327-27332	DT	denotes	these
T14901	27333-27337	NN	denotes	cell
T14900	27338-27343	NNS	denotes	lines
T14902	27344-27348	WDT	denotes	that
T14903	27349-27352	VBP	denotes	are
T14904	27353-27359	JJ	denotes	absent
T14905	27360-27362	IN	denotes	in
T14906	27363-27368	JJ	denotes	early
T14908	27369-27378	JJ	denotes	embryonic
T14907	27379-27384	NNS	denotes	cells
T14909	27384-27385	.	denotes	.
T14910	27385-27630	sentence	denotes	The repeated failure to achieve gene conversion of Capn2+/- to Capn2-/- ES cells by selection of clones in high concentrations of G418 suggests that the homozygous mutant state somehow compromises cell viability or clonal selection of ES cells.
T14911	27386-27389	DT	denotes	The
T14913	27390-27398	VBN	denotes	repeated
T14912	27399-27406	NN	denotes	failure
T14915	27407-27409	TO	denotes	to
T14916	27410-27417	VB	denotes	achieve
T14917	27418-27422	NN	denotes	gene
T14918	27423-27433	NN	denotes	conversion
T14919	27434-27436	IN	denotes	of
T14920	27437-27442	NN	denotes	Capn2
T14921	27442-27443	SYM	denotes	+
T14922	27443-27444	HYPH	denotes	/
T14923	27444-27445	SYM	denotes	-
T14924	27446-27448	IN	denotes	to
T14925	27449-27454	NN	denotes	Capn2
T14926	27454-27455	SYM	denotes	-
T14927	27455-27456	HYPH	denotes	/
T14928	27456-27457	SYM	denotes	-
T14929	27458-27460	NN	denotes	ES
T14930	27461-27466	NNS	denotes	cells
T14931	27467-27469	IN	denotes	by
T14932	27470-27479	NN	denotes	selection
T14933	27480-27482	IN	denotes	of
T14934	27483-27489	NNS	denotes	clones
T14935	27490-27492	IN	denotes	in
T14936	27493-27497	JJ	denotes	high
T14937	27498-27512	NNS	denotes	concentrations
T14938	27513-27515	IN	denotes	of
T14939	27516-27520	NN	denotes	G418
T14914	27521-27529	VBZ	denotes	suggests
T14940	27530-27534	IN	denotes	that
T14942	27535-27538	DT	denotes	the
T14944	27539-27549	JJ	denotes	homozygous
T14945	27550-27556	NN	denotes	mutant
T14943	27557-27562	NN	denotes	state
T14946	27563-27570	RB	denotes	somehow
T14941	27571-27582	VBZ	denotes	compromises
T14947	27583-27587	NN	denotes	cell
T14948	27588-27597	NN	denotes	viability
T14949	27598-27600	CC	denotes	or
T14950	27601-27607	JJ	denotes	clonal
T14951	27608-27617	NN	denotes	selection
T14952	27618-27620	IN	denotes	of
T14953	27621-27623	NN	denotes	ES
T14954	27624-27629	NNS	denotes	cells
T14955	27629-27630	.	denotes	.
T14956	27630-27851	sentence	denotes	Efforts are under way to express the m-80 k subunit from a Capn2 cDNA rescue transgene prior to gene conversion in order to preemptively rescue m-calpain activity before the remaining endogenous wild-type allele is lost.
T14957	27631-27638	NNS	denotes	Efforts
T14958	27639-27642	VBP	denotes	are
T14959	27643-27648	RB	denotes	under
T14960	27649-27652	RB	denotes	way
T14961	27653-27655	TO	denotes	to
T14962	27656-27663	VB	denotes	express
T14963	27664-27667	DT	denotes	the
T14965	27668-27669	NN	denotes	m
T14966	27669-27670	HYPH	denotes	-
T14967	27670-27672	CD	denotes	80
T14968	27673-27674	NN	denotes	k
T14964	27675-27682	NN	denotes	subunit
T14969	27683-27687	IN	denotes	from
T14970	27688-27689	DT	denotes	a
T14972	27690-27695	NN	denotes	Capn2
T14973	27696-27700	NN	denotes	cDNA
T14974	27701-27707	NN	denotes	rescue
T14971	27708-27717	NN	denotes	transgene
T14975	27718-27723	JJ	denotes	prior
T14976	27724-27726	IN	denotes	to
T14977	27727-27731	NN	denotes	gene
T14978	27732-27742	NN	denotes	conversion
T14979	27743-27745	IN	denotes	in
T14980	27746-27751	NN	denotes	order
T14981	27752-27754	TO	denotes	to
T14983	27755-27767	RB	denotes	preemptively
T14982	27768-27774	VB	denotes	rescue
T14984	27775-27776	NN	denotes	m
T14986	27776-27777	HYPH	denotes	-
T14985	27777-27784	NN	denotes	calpain
T14987	27785-27793	NN	denotes	activity
T14988	27794-27800	IN	denotes	before
T14990	27801-27804	DT	denotes	the
T14992	27805-27814	VBG	denotes	remaining
T14993	27815-27825	JJ	denotes	endogenous
T14994	27826-27830	JJ	denotes	wild
T14996	27830-27831	HYPH	denotes	-
T14995	27831-27835	NN	denotes	type
T14991	27836-27842	NN	denotes	allele
T14997	27843-27845	VBZ	denotes	is
T14989	27846-27850	VBN	denotes	lost
T14998	27850-27851	.	denotes	.
T14999	27851-27964	sentence	denotes	However, difficulty in achieving stable expression of the rescue transgene has thus far hampered these attempts.
T15000	27852-27859	RB	denotes	However
T15002	27859-27861	,	denotes	,
T15003	27861-27871	NN	denotes	difficulty
T15004	27872-27874	IN	denotes	in
T15005	27875-27884	VBG	denotes	achieving
T15006	27885-27891	JJ	denotes	stable
T15007	27892-27902	NN	denotes	expression
T15008	27903-27905	IN	denotes	of
T15009	27906-27909	DT	denotes	the
T15011	27910-27916	NN	denotes	rescue
T15010	27917-27926	NN	denotes	transgene
T15012	27927-27930	VBZ	denotes	has
T15013	27931-27935	RB	denotes	thus
T15014	27936-27939	RB	denotes	far
T15001	27940-27948	VBN	denotes	hampered
T15015	27949-27954	DT	denotes	these
T15016	27955-27963	NNS	denotes	attempts
T15017	27963-27964	.	denotes	.
T15018	27964-28144	sentence	denotes	One curiosity which arose during the genotyping of progeny from heterozygous interbreeding was the highly non-Mendelian ratio of Capn2+/+ (11.6%) to Capn2+/- (88.4%) of weanlings.
T15019	27965-27968	CD	denotes	One
T15020	27969-27978	NN	denotes	curiosity
T15022	27979-27984	WDT	denotes	which
T15023	27985-27990	VBD	denotes	arose
T15024	27991-27997	IN	denotes	during
T15025	27998-28001	DT	denotes	the
T15026	28002-28012	NN	denotes	genotyping
T15027	28013-28015	IN	denotes	of
T15028	28016-28023	NN	denotes	progeny
T15029	28024-28028	IN	denotes	from
T15030	28029-28041	JJ	denotes	heterozygous
T15031	28042-28055	NN	denotes	interbreeding
T15021	28056-28059	VBD	denotes	was
T15032	28060-28063	DT	denotes	the
T15034	28064-28070	RB	denotes	highly
T15035	28071-28084	JJ	denotes	non-Mendelian
T15033	28085-28090	NN	denotes	ratio
T15036	28091-28093	IN	denotes	of
T15037	28094-28099	NN	denotes	Capn2
T15038	28099-28100	SYM	denotes	+
T15039	28100-28101	HYPH	denotes	/
T15040	28101-28102	SYM	denotes	+
T15041	28103-28104	-LRB-	denotes	(
T15043	28104-28108	CD	denotes	11.6
T15042	28108-28109	NN	denotes	%
T15044	28109-28110	-RRB-	denotes	)
T15045	28111-28113	IN	denotes	to
T15046	28114-28119	NN	denotes	Capn2
T15047	28119-28120	SYM	denotes	+
T15048	28120-28121	HYPH	denotes	/
T15049	28121-28122	SYM	denotes	-
T15050	28123-28124	-LRB-	denotes	(
T15052	28124-28128	CD	denotes	88.4
T15051	28128-28129	NN	denotes	%
T15053	28129-28130	-RRB-	denotes	)
T15054	28131-28133	IN	denotes	of
T15055	28134-28143	NNS	denotes	weanlings
T15056	28143-28144	.	denotes	.
T15057	28144-28205	sentence	denotes	At present, there is no obvious explanation for this result.
T15058	28145-28147	IN	denotes	At
T15060	28148-28155	JJ	denotes	present
T15061	28155-28157	,	denotes	,
T15062	28157-28162	EX	denotes	there
T15059	28163-28165	VBZ	denotes	is
T15063	28166-28168	DT	denotes	no
T15065	28169-28176	JJ	denotes	obvious
T15064	28177-28188	NN	denotes	explanation
T15066	28189-28192	IN	denotes	for
T15067	28193-28197	DT	denotes	this
T15068	28198-28204	NN	denotes	result
T15069	28204-28205	.	denotes	.
T15070	28205-28342	sentence	denotes	Crosses between wild-type and heterozygous mice also produced progeny with a greater than expected proportion of heterozygous offspring.
T15071	28206-28213	NNS	denotes	Crosses
T15073	28214-28221	IN	denotes	between
T15074	28222-28226	JJ	denotes	wild
T15076	28226-28227	HYPH	denotes	-
T15075	28227-28231	NN	denotes	type
T15078	28232-28235	CC	denotes	and
T15079	28236-28248	JJ	denotes	heterozygous
T15077	28249-28253	NNS	denotes	mice
T15080	28254-28258	RB	denotes	also
T15072	28259-28267	VBD	denotes	produced
T15081	28268-28275	NN	denotes	progeny
T15082	28276-28280	IN	denotes	with
T15083	28281-28282	DT	denotes	a
T15085	28283-28290	JJR	denotes	greater
T15086	28291-28295	IN	denotes	than
T15087	28296-28304	VBN	denotes	expected
T15084	28305-28315	NN	denotes	proportion
T15088	28316-28318	IN	denotes	of
T15089	28319-28331	JJ	denotes	heterozygous
T15090	28332-28341	NN	denotes	offspring
T15091	28341-28342	.	denotes	.
T15092	28342-28548	sentence	denotes	Interestingly, heterozygous crosses involving the Capn4 transgenic line generated by Zimmerman and colleagues yielded progeny with a similar, if less extreme, skewing in favor of the heterozygous genotype.
T15093	28343-28356	RB	denotes	Interestingly
T15095	28356-28358	,	denotes	,
T15096	28358-28370	JJ	denotes	heterozygous
T15097	28371-28378	NNS	denotes	crosses
T15098	28379-28388	VBG	denotes	involving
T15099	28389-28392	DT	denotes	the
T15101	28393-28398	NN	denotes	Capn4
T15102	28399-28409	JJ	denotes	transgenic
T15100	28410-28414	NN	denotes	line
T15103	28415-28424	VBN	denotes	generated
T15104	28425-28427	IN	denotes	by
T15105	28428-28437	NNP	denotes	Zimmerman
T15106	28438-28441	CC	denotes	and
T15107	28442-28452	NNS	denotes	colleagues
T15094	28453-28460	VBD	denotes	yielded
T15108	28461-28468	NN	denotes	progeny
T15109	28469-28473	IN	denotes	with
T15110	28474-28475	DT	denotes	a
T15112	28476-28483	JJ	denotes	similar
T15113	28483-28485	,	denotes	,
T15115	28485-28487	IN	denotes	if
T15116	28488-28492	RBR	denotes	less
T15114	28493-28500	JJ	denotes	extreme
T15117	28500-28502	,	denotes	,
T15111	28502-28509	VBG	denotes	skewing
T15118	28510-28512	IN	denotes	in
T15119	28513-28518	NN	denotes	favor
T15120	28519-28521	IN	denotes	of
T15121	28522-28525	DT	denotes	the
T15123	28526-28538	JJ	denotes	heterozygous
T15122	28539-28547	NN	denotes	genotype
T15124	28547-28548	.	denotes	.
T15125	28548-28685	sentence	denotes	Out of a total of 80 genotyped animals, 22.5% were wild-type and 77.5% were heterozygous, with no homozygous null progeny observed [32].
T15126	28549-28552	IN	denotes	Out
T15128	28553-28555	IN	denotes	of
T15129	28556-28557	DT	denotes	a
T15130	28558-28563	NN	denotes	total
T15131	28564-28566	IN	denotes	of
T15132	28567-28569	CD	denotes	80
T15134	28570-28579	VBN	denotes	genotyped
T15133	28580-28587	NNS	denotes	animals
T15135	28587-28589	,	denotes	,
T15136	28589-28593	CD	denotes	22.5
T15137	28593-28594	NN	denotes	%
T15127	28595-28599	VBD	denotes	were
T15138	28600-28604	JJ	denotes	wild
T15140	28604-28605	HYPH	denotes	-
T15139	28605-28609	NN	denotes	type
T15141	28610-28613	CC	denotes	and
T15142	28614-28618	CD	denotes	77.5
T15143	28618-28619	NN	denotes	%
T15144	28620-28624	VBD	denotes	were
T15145	28625-28637	JJ	denotes	heterozygous
T15146	28637-28639	,	denotes	,
T15147	28639-28643	IN	denotes	with
T15149	28644-28646	DT	denotes	no
T15151	28647-28657	JJ	denotes	homozygous
T15152	28658-28662	JJ	denotes	null
T15150	28663-28670	NN	denotes	progeny
T15148	28671-28679	VBN	denotes	observed
T15153	28680-28681	-LRB-	denotes	[
T15154	28681-28683	CD	denotes	32
T15155	28683-28684	-RRB-	denotes	]
T15156	28684-28685	.	denotes	.
T15157	28685-28782	sentence	denotes	These observations suggest a developmental advantage associated with reduced calpain expression.
T15158	28686-28691	DT	denotes	These
T15159	28692-28704	NNS	denotes	observations
T15160	28705-28712	VBP	denotes	suggest
T15161	28713-28714	DT	denotes	a
T15163	28715-28728	JJ	denotes	developmental
T15162	28729-28738	NN	denotes	advantage
T15164	28739-28749	VBN	denotes	associated
T15165	28750-28754	IN	denotes	with
T15166	28755-28762	VBN	denotes	reduced
T15168	28763-28770	NN	denotes	calpain
T15167	28771-28781	NN	denotes	expression
T15169	28781-28782	.	denotes	.
T15170	28782-28847	sentence	denotes	Perhaps future studies will reveal a mechanistic basis for this.
T15171	28783-28790	RB	denotes	Perhaps
T15173	28791-28797	JJ	denotes	future
T15174	28798-28805	NNS	denotes	studies
T15175	28806-28810	MD	denotes	will
T15172	28811-28817	VB	denotes	reveal
T15176	28818-28819	DT	denotes	a
T15178	28820-28831	JJ	denotes	mechanistic
T15177	28832-28837	NN	denotes	basis
T15179	28838-28841	IN	denotes	for
T15180	28842-28846	DT	denotes	this
T15181	28846-28847	.	denotes	.
T15430	28860-28863	DT	denotes	The
T15431	28864-28868	NN	denotes	work
T15433	28869-28878	VBN	denotes	presented
T15434	28879-28883	RB	denotes	here
T15435	28884-28887	VBZ	denotes	has
T15432	28888-28897	VBN	denotes	clarified
T15436	28898-28901	CD	denotes	two
T15438	28902-28911	JJ	denotes	important
T15437	28912-28921	NNS	denotes	questions
T15439	28922-28931	VBG	denotes	regarding
T15440	28932-28935	DT	denotes	the
T15442	28936-28949	JJ	denotes	physiological
T15441	28950-28955	NNS	denotes	roles
T15443	28956-28958	IN	denotes	of
T15444	28959-28962	DT	denotes	the
T15446	28963-28966	CD	denotes	two
T15447	28967-28977	JJ	denotes	ubiquitous
T15448	28978-28985	NN	denotes	calpain
T15445	28986-28994	NNS	denotes	isoforms
T15449	28994-28996	,	denotes	,
T15450	28996-28997	NN	denotes	μ
T15452	28997-28998	HYPH	denotes	-
T15453	28999-29002	CC	denotes	and
T15454	29003-29004	NN	denotes	m
T15455	29004-29005	HYPH	denotes	-
T15451	29005-29012	NN	denotes	calpain
T15456	29012-29013	.	denotes	.
T15457	29013-29158	sentence	denotes	Firstly, it was determined that m-calpain plays an indispensable role in murine embryogenesis, possibly related to pre-implantation development.
T15458	29014-29021	RB	denotes	Firstly
T15460	29021-29023	,	denotes	,
T15461	29023-29025	PRP	denotes	it
T15462	29026-29029	VBD	denotes	was
T15459	29030-29040	VBN	denotes	determined
T15463	29041-29045	IN	denotes	that
T15465	29046-29047	NN	denotes	m
T15467	29047-29048	HYPH	denotes	-
T15466	29048-29055	NN	denotes	calpain
T15464	29056-29061	VBZ	denotes	plays
T15468	29062-29064	DT	denotes	an
T15470	29065-29078	JJ	denotes	indispensable
T15469	29079-29083	NN	denotes	role
T15471	29084-29086	IN	denotes	in
T15472	29087-29093	JJ	denotes	murine
T15473	29094-29107	NN	denotes	embryogenesis
T15474	29107-29109	,	denotes	,
T15475	29109-29117	RB	denotes	possibly
T15476	29118-29125	VBN	denotes	related
T15477	29126-29128	IN	denotes	to
T15478	29129-29145	JJ	denotes	pre-implantation
T15479	29146-29157	NN	denotes	development
T15480	29157-29158	.	denotes	.
T15481	29158-29362	sentence	denotes	Furthermore, this function cannot be carried out by μ-calpain despite the apparent in vitro similarities of μ- and m-calpain, demonstrating that the two isoforms clearly have some distinct roles in vivo.
T15482	29159-29170	RB	denotes	Furthermore
T15484	29170-29172	,	denotes	,
T15485	29172-29176	DT	denotes	this
T15486	29177-29185	NN	denotes	function
T15487	29186-29189	MD	denotes	can
T15488	29189-29192	RB	denotes	not
T15489	29193-29195	VB	denotes	be
T15483	29196-29203	VBN	denotes	carried
T15490	29204-29207	RP	denotes	out
T15491	29208-29210	IN	denotes	by
T15492	29211-29212	NN	denotes	μ
T15494	29212-29213	HYPH	denotes	-
T15493	29213-29220	NN	denotes	calpain
T15495	29221-29228	IN	denotes	despite
T15496	29229-29232	DT	denotes	the
T15498	29233-29241	JJ	denotes	apparent
T15499	29242-29244	FW	denotes	in
T15500	29245-29250	FW	denotes	vitro
T15497	29251-29263	NNS	denotes	similarities
T15501	29264-29266	IN	denotes	of
T15502	29267-29268	NN	denotes	μ
T15504	29268-29269	HYPH	denotes	-
T15505	29270-29273	CC	denotes	and
T15506	29274-29275	NN	denotes	m
T15507	29275-29276	HYPH	denotes	-
T15503	29276-29283	NN	denotes	calpain
T15508	29283-29285	,	denotes	,
T15509	29285-29298	VBG	denotes	demonstrating
T15510	29299-29303	IN	denotes	that
T15512	29304-29307	DT	denotes	the
T15514	29308-29311	CD	denotes	two
T15513	29312-29320	NNS	denotes	isoforms
T15515	29321-29328	RB	denotes	clearly
T15511	29329-29333	VBP	denotes	have
T15516	29334-29338	DT	denotes	some
T15518	29339-29347	JJ	denotes	distinct
T15517	29348-29353	NNS	denotes	roles
T15519	29354-29356	FW	denotes	in
T15520	29357-29361	FW	denotes	vivo
T15521	29361-29362	.	denotes	.
T15522	29362-29545	sentence	denotes	The functions of m-calpain during post-implantation embryogenesis and in adult mice remain to be elucidated and will have to be addressed using a conditional gene targeting strategy.
T15523	29363-29366	DT	denotes	The
T15524	29367-29376	NNS	denotes	functions
T15526	29377-29379	IN	denotes	of
T15527	29380-29381	NN	denotes	m
T15529	29381-29382	HYPH	denotes	-
T15528	29382-29389	NN	denotes	calpain
T15530	29390-29396	IN	denotes	during
T15531	29397-29414	JJ	denotes	post-implantation
T15532	29415-29428	NN	denotes	embryogenesis
T15533	29429-29432	CC	denotes	and
T15534	29433-29435	IN	denotes	in
T15535	29436-29441	JJ	denotes	adult
T15536	29442-29446	NNS	denotes	mice
T15525	29447-29453	VBP	denotes	remain
T15537	29454-29456	TO	denotes	to
T15539	29457-29459	VB	denotes	be
T15538	29460-29470	VBN	denotes	elucidated
T15540	29471-29474	CC	denotes	and
T15541	29475-29479	MD	denotes	will
T15542	29480-29484	VB	denotes	have
T15543	29485-29487	TO	denotes	to
T15545	29488-29490	VB	denotes	be
T15544	29491-29500	VBN	denotes	addressed
T15546	29501-29506	VBG	denotes	using
T15547	29507-29508	DT	denotes	a
T15549	29509-29520	JJ	denotes	conditional
T15550	29521-29525	NN	denotes	gene
T15551	29526-29535	NN	denotes	targeting
T15548	29536-29544	NN	denotes	strategy
T15552	29544-29545	.	denotes	.
T16008	29556-29563	NN	denotes	Cloning
T16009	29564-29567	CC	denotes	and
T16010	29568-29578	NN	denotes	sequencing
T16011	29579-29581	IN	denotes	of
T16012	29582-29585	DT	denotes	the
T16014	29586-29591	NN	denotes	mouse
T16015	29592-29597	NN	denotes	Capn2
T16013	29598-29603	NN	denotes	locus
T16016	29603-29755	sentence	denotes	The mouse Capn2 gene encodes the 700 amino acids of the m-calpain large subunit (m-80 k) and consists of 21 exons extending over 50-kb on chromosome 1.
T16017	29604-29607	DT	denotes	The
T16019	29608-29613	NN	denotes	mouse
T16020	29614-29619	NN	denotes	Capn2
T16018	29620-29624	NN	denotes	gene
T16021	29625-29632	VBZ	denotes	encodes
T16022	29633-29636	DT	denotes	the
T16024	29637-29640	CD	denotes	700
T16025	29641-29646	NN	denotes	amino
T16023	29647-29652	NNS	denotes	acids
T16026	29653-29655	IN	denotes	of
T16027	29656-29659	DT	denotes	the
T16029	29660-29661	NN	denotes	m
T16031	29661-29662	HYPH	denotes	-
T16030	29662-29669	NN	denotes	calpain
T16032	29670-29675	JJ	denotes	large
T16028	29676-29683	NN	denotes	subunit
T16033	29684-29685	-LRB-	denotes	(
T16035	29685-29686	NN	denotes	m
T16036	29686-29687	HYPH	denotes	-
T16037	29687-29689	CD	denotes	80
T16034	29690-29691	NN	denotes	k
T16038	29691-29692	-RRB-	denotes	)
T16039	29693-29696	CC	denotes	and
T16040	29697-29705	VBZ	denotes	consists
T16041	29706-29708	IN	denotes	of
T16042	29709-29711	CD	denotes	21
T16043	29712-29717	NNS	denotes	exons
T16044	29718-29727	VBG	denotes	extending
T16045	29728-29732	IN	denotes	over
T16046	29733-29735	CD	denotes	50
T16048	29735-29736	HYPH	denotes	-
T16047	29736-29738	NN	denotes	kb
T16049	29739-29741	IN	denotes	on
T16050	29742-29752	NN	denotes	chromosome
T16051	29753-29754	CD	denotes	1
T16052	29754-29755	.	denotes	.
T16053	29755-29887	sentence	denotes	A cDNA clone encoding a portion of the mouse m-80 k subunit was purchased (dbEST Id, 807416, Image:606689, Image Consortium, LLNL).
T16054	29756-29757	DT	denotes	A
T16056	29758-29762	NN	denotes	cDNA
T16055	29763-29768	NN	denotes	clone
T16058	29769-29777	VBG	denotes	encoding
T16059	29778-29779	DT	denotes	a
T16060	29780-29787	NN	denotes	portion
T16061	29788-29790	IN	denotes	of
T16062	29791-29794	DT	denotes	the
T16064	29795-29800	NN	denotes	mouse
T16065	29801-29802	NN	denotes	m
T16067	29802-29803	HYPH	denotes	-
T16068	29803-29805	CD	denotes	80
T16066	29806-29807	NN	denotes	k
T16063	29808-29815	NN	denotes	subunit
T16069	29816-29819	VBD	denotes	was
T16057	29820-29829	VBN	denotes	purchased
T16070	29830-29831	-LRB-	denotes	(
T16072	29831-29836	NN	denotes	dbEST
T16073	29837-29839	NN	denotes	Id
T16074	29839-29841	,	denotes	,
T16075	29841-29847	CD	denotes	807416
T16076	29847-29849	,	denotes	,
T16077	29849-29854	NNP	denotes	Image
T16078	29854-29855	:	denotes	:
T16079	29855-29861	CD	denotes	606689
T16080	29861-29863	,	denotes	,
T16081	29863-29868	NNP	denotes	Image
T16082	29869-29879	NNP	denotes	Consortium
T16083	29879-29881	,	denotes	,
T16071	29881-29885	NNP	denotes	LLNL
T16084	29885-29886	-RRB-	denotes	)
T16085	29886-29887	.	denotes	.
T16086	29887-30050	sentence	denotes	It was found to contain 2.8-kb of sequence from position 247 of the coding sequence to the stop codon at position 2,101, including the 3'-UTR to the polyA signal.
T16087	29888-29890	PRP	denotes	It
T16089	29891-29894	VBD	denotes	was
T16088	29895-29900	VBN	denotes	found
T16090	29901-29903	TO	denotes	to
T16091	29904-29911	VB	denotes	contain
T16092	29912-29915	CD	denotes	2.8
T16094	29915-29916	HYPH	denotes	-
T16093	29916-29918	NN	denotes	kb
T16095	29919-29921	IN	denotes	of
T16096	29922-29930	NN	denotes	sequence
T16097	29931-29935	IN	denotes	from
T16098	29936-29944	NN	denotes	position
T16099	29945-29948	CD	denotes	247
T16100	29949-29951	IN	denotes	of
T16101	29952-29955	DT	denotes	the
T16103	29956-29962	NN	denotes	coding
T16102	29963-29971	NN	denotes	sequence
T16104	29972-29974	IN	denotes	to
T16105	29975-29978	DT	denotes	the
T16107	29979-29983	NN	denotes	stop
T16106	29984-29989	NN	denotes	codon
T16108	29990-29992	IN	denotes	at
T16109	29993-30001	NN	denotes	position
T16110	30002-30007	CD	denotes	2,101
T16111	30007-30009	,	denotes	,
T16112	30009-30018	IN	denotes	including
T16113	30019-30022	DT	denotes	the
T16115	30023-30024	CD	denotes	3
T16116	30024-30025	SYM	denotes	'
T16117	30025-30026	HYPH	denotes	-
T16114	30026-30029	NN	denotes	UTR
T16118	30030-30032	IN	denotes	to
T16119	30033-30036	DT	denotes	the
T16121	30037-30042	NN	denotes	polyA
T16120	30043-30049	NN	denotes	signal
T16122	30049-30050	.	denotes	.
T16123	30050-30151	sentence	denotes	A fragment of this cDNA was subcloned and used to screen a 129Sv mouse genomic library in λ-Dash II.
T16124	30051-30052	DT	denotes	A
T16125	30053-30061	NN	denotes	fragment
T16127	30062-30064	IN	denotes	of
T16128	30065-30069	DT	denotes	this
T16129	30070-30074	NN	denotes	cDNA
T16130	30075-30078	VBD	denotes	was
T16126	30079-30088	VBN	denotes	subcloned
T16131	30089-30092	CC	denotes	and
T16132	30093-30097	VBN	denotes	used
T16133	30098-30100	TO	denotes	to
T16134	30101-30107	VB	denotes	screen
T16135	30108-30109	DT	denotes	a
T16137	30110-30115	CD	denotes	129Sv
T16138	30116-30121	NN	denotes	mouse
T16139	30122-30129	JJ	denotes	genomic
T16136	30130-30137	NN	denotes	library
T16140	30138-30140	IN	denotes	in
T16141	30141-30142	NN	denotes	λ
T16143	30142-30143	HYPH	denotes	-
T16142	30143-30147	NN	denotes	Dash
T16144	30148-30150	CD	denotes	II
T16145	30150-30151	.	denotes	.
T16146	30151-30282	sentence	denotes	Overlapping genomic clones were obtained covering 15,354 bp, extending from a BamHI site in intron 3 to a BamHI site in intron 15.
T16147	30152-30163	VBG	denotes	Overlapping
T16149	30164-30171	JJ	denotes	genomic
T16148	30172-30178	NNS	denotes	clones
T16151	30179-30183	VBD	denotes	were
T16150	30184-30192	VBN	denotes	obtained
T16152	30193-30201	VBG	denotes	covering
T16153	30202-30208	CD	denotes	15,354
T16154	30209-30211	NN	denotes	bp
T16155	30211-30213	,	denotes	,
T16156	30213-30222	VBG	denotes	extending
T16157	30223-30227	IN	denotes	from
T16158	30228-30229	DT	denotes	a
T16160	30230-30235	NN	denotes	BamHI
T16159	30236-30240	NN	denotes	site
T16161	30241-30243	IN	denotes	in
T16162	30244-30250	NN	denotes	intron
T16163	30251-30252	CD	denotes	3
T16164	30253-30255	IN	denotes	to
T16165	30256-30257	DT	denotes	a
T16167	30258-30263	NN	denotes	BamHI
T16166	30264-30268	NN	denotes	site
T16168	30269-30271	IN	denotes	in
T16169	30272-30278	NN	denotes	intron
T16170	30279-30281	CD	denotes	15
T16171	30281-30282	.	denotes	.
T16172	30282-30402	sentence	denotes	These clones were sequenced completely on both strands and the data were submitted to GenBank (accession no. AF497625).
T16173	30283-30288	DT	denotes	These
T16174	30289-30295	NNS	denotes	clones
T16176	30296-30300	VBD	denotes	were
T16175	30301-30310	VBN	denotes	sequenced
T16177	30311-30321	RB	denotes	completely
T16178	30322-30324	IN	denotes	on
T16179	30325-30329	DT	denotes	both
T16180	30330-30337	NNS	denotes	strands
T16181	30338-30341	CC	denotes	and
T16182	30342-30345	DT	denotes	the
T16183	30346-30350	NNS	denotes	data
T16185	30351-30355	VBD	denotes	were
T16184	30356-30365	VBN	denotes	submitted
T16186	30366-30368	IN	denotes	to
T16187	30369-30376	NNP	denotes	GenBank
T16188	30377-30378	-LRB-	denotes	(
T16190	30378-30387	NN	denotes	accession
T16191	30388-30391	NN	denotes	no.
T16189	30392-30400	NN	denotes	AF497625
T16192	30400-30401	-RRB-	denotes	)
T16193	30401-30402	.	denotes	.
T16194	30402-30642	sentence	denotes	The submitted sequences agreed precisely with the public databases, and also filled in several small gaps corresponding to short repetitive sequences which could only be firmly established by repeated sequencing in non-standard conditions.
T16195	30403-30406	DT	denotes	The
T16197	30407-30416	VBN	denotes	submitted
T16196	30417-30426	NNS	denotes	sequences
T16198	30427-30433	VBD	denotes	agreed
T16199	30434-30443	RB	denotes	precisely
T16200	30444-30448	IN	denotes	with
T16201	30449-30452	DT	denotes	the
T16203	30453-30459	JJ	denotes	public
T16202	30460-30469	NNS	denotes	databases
T16204	30469-30471	,	denotes	,
T16205	30471-30474	CC	denotes	and
T16206	30475-30479	RB	denotes	also
T16207	30480-30486	VBN	denotes	filled
T16208	30487-30489	RP	denotes	in
T16209	30490-30497	JJ	denotes	several
T16211	30498-30503	JJ	denotes	small
T16210	30504-30508	NNS	denotes	gaps
T16212	30509-30522	VBG	denotes	corresponding
T16213	30523-30525	IN	denotes	to
T16214	30526-30531	JJ	denotes	short
T16216	30532-30542	JJ	denotes	repetitive
T16215	30543-30552	NNS	denotes	sequences
T16217	30553-30558	WDT	denotes	which
T16219	30559-30564	MD	denotes	could
T16220	30565-30569	RB	denotes	only
T16221	30570-30572	VB	denotes	be
T16222	30573-30579	RB	denotes	firmly
T16218	30580-30591	VBN	denotes	established
T16223	30592-30594	IN	denotes	by
T16224	30595-30603	VBN	denotes	repeated
T16225	30604-30614	NN	denotes	sequencing
T16226	30615-30617	IN	denotes	in
T16227	30618-30630	JJ	denotes	non-standard
T16228	30631-30641	NNS	denotes	conditions
T16229	30641-30642	.	denotes	.
T16709	30644-30656	NN	denotes	Construction
T16710	30657-30659	IN	denotes	of
T16711	30660-30663	DT	denotes	the
T16713	30664-30669	NN	denotes	Capn2
T16714	30670-30679	NN	denotes	targeting
T16712	30680-30686	NN	denotes	vector
T16715	30686-30840	sentence	denotes	A targeting construct was designed to replace a 785 bp BamHI-HindIII fragment, containing exon 7 of the Capn2 gene, with the PGK-Neo cassette (Figure 1).
T16716	30687-30688	DT	denotes	A
T16718	30689-30698	NN	denotes	targeting
T16717	30699-30708	NN	denotes	construct
T16720	30709-30712	VBD	denotes	was
T16719	30713-30721	VBN	denotes	designed
T16721	30722-30724	TO	denotes	to
T16722	30725-30732	VB	denotes	replace
T16723	30733-30734	DT	denotes	a
T16725	30735-30738	CD	denotes	785
T16726	30739-30741	NN	denotes	bp
T16727	30742-30747	NN	denotes	BamHI
T16729	30747-30748	HYPH	denotes	-
T16728	30748-30755	NN	denotes	HindIII
T16724	30756-30764	NN	denotes	fragment
T16730	30764-30766	,	denotes	,
T16731	30766-30776	VBG	denotes	containing
T16732	30777-30781	NN	denotes	exon
T16733	30782-30783	CD	denotes	7
T16734	30784-30786	IN	denotes	of
T16735	30787-30790	DT	denotes	the
T16737	30791-30796	NN	denotes	Capn2
T16736	30797-30801	NN	denotes	gene
T16738	30801-30803	,	denotes	,
T16739	30803-30807	IN	denotes	with
T16740	30808-30811	DT	denotes	the
T16742	30812-30815	NN	denotes	PGK
T16744	30815-30816	HYPH	denotes	-
T16743	30816-30819	NN	denotes	Neo
T16741	30820-30828	NN	denotes	cassette
T16745	30829-30830	-LRB-	denotes	(
T16746	30830-30836	NN	denotes	Figure
T16747	30837-30838	CD	denotes	1
T16748	30838-30839	-RRB-	denotes	)
T16749	30839-30840	.	denotes	.
T16750	30840-30952	sentence	denotes	Exon 7 encodes 24 amino acids in the active-site region, including Asn286, one of the catalytic triad residues.
T16751	30841-30845	NN	denotes	Exon
T16753	30846-30847	CD	denotes	7
T16752	30848-30855	VBZ	denotes	encodes
T16754	30856-30858	CD	denotes	24
T16756	30859-30864	NN	denotes	amino
T16755	30865-30870	NNS	denotes	acids
T16757	30871-30873	IN	denotes	in
T16758	30874-30877	DT	denotes	the
T16760	30878-30884	JJ	denotes	active
T16762	30884-30885	HYPH	denotes	-
T16761	30885-30889	NN	denotes	site
T16759	30890-30896	NN	denotes	region
T16763	30896-30898	,	denotes	,
T16764	30898-30907	VBG	denotes	including
T16765	30908-30914	NN	denotes	Asn286
T16766	30914-30916	,	denotes	,
T16767	30916-30919	CD	denotes	one
T16768	30920-30922	IN	denotes	of
T16769	30923-30926	DT	denotes	the
T16771	30927-30936	JJ	denotes	catalytic
T16772	30937-30942	NN	denotes	triad
T16770	30943-30951	NNS	denotes	residues
T16773	30951-30952	.	denotes	.
T16774	30952-31158	sentence	denotes	The short (upstream) arm of the targeting construct was provided by a 2.7-kb HindIII-BamHI fragment, containing exons 5 and 6, which was inserted into the pPNT vector upstream of the PGK-Neo cassette [48].
T16775	30953-30956	DT	denotes	The
T16777	30957-30962	JJ	denotes	short
T16778	30963-30964	-LRB-	denotes	(
T16779	30964-30972	JJ	denotes	upstream
T16780	30972-30973	-RRB-	denotes	)
T16776	30974-30977	NN	denotes	arm
T16782	30978-30980	IN	denotes	of
T16783	30981-30984	DT	denotes	the
T16785	30985-30994	NN	denotes	targeting
T16784	30995-31004	NN	denotes	construct
T16786	31005-31008	VBD	denotes	was
T16781	31009-31017	VBN	denotes	provided
T16787	31018-31020	IN	denotes	by
T16788	31021-31022	DT	denotes	a
T16790	31023-31026	CD	denotes	2.7
T16792	31026-31027	HYPH	denotes	-
T16791	31027-31029	NN	denotes	kb
T16793	31030-31037	NN	denotes	HindIII
T16795	31037-31038	HYPH	denotes	-
T16794	31038-31043	NN	denotes	BamHI
T16789	31044-31052	NN	denotes	fragment
T16796	31052-31054	,	denotes	,
T16797	31054-31064	VBG	denotes	containing
T16798	31065-31070	NNS	denotes	exons
T16799	31071-31072	CD	denotes	5
T16800	31073-31076	CC	denotes	and
T16801	31077-31078	CD	denotes	6
T16802	31078-31080	,	denotes	,
T16803	31080-31085	WDT	denotes	which
T16805	31086-31089	VBD	denotes	was
T16804	31090-31098	VBN	denotes	inserted
T16806	31099-31103	IN	denotes	into
T16807	31104-31107	DT	denotes	the
T16809	31108-31112	NN	denotes	pPNT
T16808	31113-31119	NN	denotes	vector
T16810	31120-31128	RB	denotes	upstream
T16811	31129-31131	IN	denotes	of
T16812	31132-31135	DT	denotes	the
T16814	31136-31139	NN	denotes	PGK
T16816	31139-31140	HYPH	denotes	-
T16815	31140-31143	NN	denotes	Neo
T16813	31144-31152	NN	denotes	cassette
T16817	31153-31154	-LRB-	denotes	[
T16818	31154-31156	CD	denotes	48
T16819	31156-31157	-RRB-	denotes	]
T16820	31157-31158	.	denotes	.
T16821	31158-31235	sentence	denotes	During cloning, the BamHI site at the 3' end of the short arm was abolished.
T16822	31159-31165	IN	denotes	During
T16824	31166-31173	NN	denotes	cloning
T16825	31173-31175	,	denotes	,
T16826	31175-31178	DT	denotes	the
T16828	31179-31184	NN	denotes	BamHI
T16827	31185-31189	NN	denotes	site
T16829	31190-31192	IN	denotes	at
T16830	31193-31196	DT	denotes	the
T16832	31197-31198	CD	denotes	3
T16833	31198-31199	SYM	denotes	'
T16831	31200-31203	NN	denotes	end
T16834	31204-31206	IN	denotes	of
T16835	31207-31210	DT	denotes	the
T16837	31211-31216	JJ	denotes	short
T16836	31217-31220	NN	denotes	arm
T16838	31221-31224	VBD	denotes	was
T16823	31225-31234	VBN	denotes	abolished
T16839	31234-31235	.	denotes	.
T16840	31235-31465	sentence	denotes	The loss of this BamHI site in the mutant allele, coupled with the introduction of a new BamHI site at the 3' end of the PGK-Neo cassette, provided a basis for distinguishing the wild-type and mutant alleles by Southern blotting.
T16841	31236-31239	DT	denotes	The
T16842	31240-31244	NN	denotes	loss
T16844	31245-31247	IN	denotes	of
T16845	31248-31252	DT	denotes	this
T16847	31253-31258	NN	denotes	BamHI
T16846	31259-31263	NN	denotes	site
T16848	31264-31266	IN	denotes	in
T16849	31267-31270	DT	denotes	the
T16851	31271-31277	NN	denotes	mutant
T16850	31278-31284	NN	denotes	allele
T16852	31284-31286	,	denotes	,
T16853	31286-31293	VBN	denotes	coupled
T16854	31294-31298	IN	denotes	with
T16855	31299-31302	DT	denotes	the
T16856	31303-31315	NN	denotes	introduction
T16857	31316-31318	IN	denotes	of
T16858	31319-31320	DT	denotes	a
T16860	31321-31324	JJ	denotes	new
T16861	31325-31330	NN	denotes	BamHI
T16859	31331-31335	NN	denotes	site
T16862	31336-31338	IN	denotes	at
T16863	31339-31342	DT	denotes	the
T16865	31343-31344	CD	denotes	3
T16866	31344-31345	SYM	denotes	'
T16864	31346-31349	NN	denotes	end
T16867	31350-31352	IN	denotes	of
T16868	31353-31356	DT	denotes	the
T16870	31357-31360	NN	denotes	PGK
T16872	31360-31361	HYPH	denotes	-
T16871	31361-31364	NN	denotes	Neo
T16869	31365-31373	NN	denotes	cassette
T16873	31373-31375	,	denotes	,
T16843	31375-31383	VBN	denotes	provided
T16874	31384-31385	DT	denotes	a
T16875	31386-31391	NN	denotes	basis
T16876	31392-31395	IN	denotes	for
T16877	31396-31410	VBG	denotes	distinguishing
T16878	31411-31414	DT	denotes	the
T16880	31415-31419	JJ	denotes	wild
T16882	31419-31420	HYPH	denotes	-
T16881	31420-31424	NN	denotes	type
T16883	31425-31428	CC	denotes	and
T16884	31429-31435	NN	denotes	mutant
T16879	31436-31443	NNS	denotes	alleles
T16885	31444-31446	IN	denotes	by
T16886	31447-31455	NNP	denotes	Southern
T16887	31456-31464	NN	denotes	blotting
T16888	31464-31465	.	denotes	.
T16889	31465-31653	sentence	denotes	The long (downstream) arm of homology was provided by a 7-kb HindIII-KpnI fragment, extending from intron 7 to intron 12, inserted between the PGK-Neo and thymidine kinase (tk) cassettes.
T16890	31466-31469	DT	denotes	The
T16892	31470-31474	JJ	denotes	long
T16893	31475-31476	-LRB-	denotes	(
T16894	31476-31486	JJ	denotes	downstream
T16895	31486-31487	-RRB-	denotes	)
T16891	31488-31491	NN	denotes	arm
T16897	31492-31494	IN	denotes	of
T16898	31495-31503	NN	denotes	homology
T16899	31504-31507	VBD	denotes	was
T16896	31508-31516	VBN	denotes	provided
T16900	31517-31519	IN	denotes	by
T16901	31520-31521	DT	denotes	a
T16903	31522-31523	CD	denotes	7
T16905	31523-31524	HYPH	denotes	-
T16904	31524-31526	NN	denotes	kb
T16906	31527-31534	NN	denotes	HindIII
T16908	31534-31535	HYPH	denotes	-
T16907	31535-31539	NN	denotes	KpnI
T16902	31540-31548	NN	denotes	fragment
T16909	31548-31550	,	denotes	,
T16910	31550-31559	VBG	denotes	extending
T16911	31560-31564	IN	denotes	from
T16912	31565-31571	NN	denotes	intron
T16913	31572-31573	CD	denotes	7
T16914	31574-31576	IN	denotes	to
T16915	31577-31583	NN	denotes	intron
T16916	31584-31586	CD	denotes	12
T16917	31586-31588	,	denotes	,
T16918	31588-31596	VBN	denotes	inserted
T16919	31597-31604	IN	denotes	between
T16920	31605-31608	DT	denotes	the
T16922	31609-31612	NN	denotes	PGK
T16924	31612-31613	HYPH	denotes	-
T16923	31613-31616	NN	denotes	Neo
T16925	31617-31620	CC	denotes	and
T16926	31621-31630	NN	denotes	thymidine
T16927	31631-31637	NN	denotes	kinase
T16928	31638-31639	-LRB-	denotes	(
T16929	31639-31641	NN	denotes	tk
T16930	31641-31642	-RRB-	denotes	)
T16921	31643-31652	NNS	denotes	cassettes
T16931	31652-31653	.	denotes	.
T23423	31664-31673	NN	denotes	Targeting
T23424	31674-31682	NN	denotes	strategy
T23425	31683-31686	IN	denotes	for
T23426	31687-31697	NN	denotes	disruption
T23427	31698-31700	IN	denotes	of
T23428	31701-31704	DT	denotes	the
T23430	31705-31711	JJ	denotes	murine
T23431	31712-31717	NN	denotes	Capn2
T23429	31718-31722	NN	denotes	gene
T23432	31722-31723	.	denotes	.
T23433	31723-31830	sentence	denotes	The murine Capn2 gene, encoding the m-80 k subunit, was disrupted in ES cells by homologous recombination.
T23434	31724-31727	DT	denotes	The
T23436	31728-31734	JJ	denotes	murine
T23437	31735-31740	NN	denotes	Capn2
T23435	31741-31745	NN	denotes	gene
T23439	31745-31747	,	denotes	,
T23440	31747-31755	VBG	denotes	encoding
T23441	31756-31759	DT	denotes	the
T23443	31760-31761	NN	denotes	m
T23445	31761-31762	HYPH	denotes	-
T23446	31762-31764	CD	denotes	80
T23444	31765-31766	NN	denotes	k
T23442	31767-31774	NN	denotes	subunit
T23447	31774-31776	,	denotes	,
T23448	31776-31779	VBD	denotes	was
T23438	31780-31789	VBN	denotes	disrupted
T23449	31790-31792	IN	denotes	in
T23450	31793-31795	NN	denotes	ES
T23451	31796-31801	NNS	denotes	cells
T23452	31802-31804	IN	denotes	by
T23453	31805-31815	JJ	denotes	homologous
T23454	31816-31829	NN	denotes	recombination
T23455	31829-31830	.	denotes	.
T23456	31830-31952	sentence	denotes	The structures of the wild-type allele (top), the targeting vector (middle), and the mutant allele (bottom) are depicted.
T23457	31831-31834	DT	denotes	The
T23458	31835-31845	NNS	denotes	structures
T23460	31846-31848	IN	denotes	of
T23461	31849-31852	DT	denotes	the
T23463	31853-31857	JJ	denotes	wild
T23465	31857-31858	HYPH	denotes	-
T23464	31858-31862	NN	denotes	type
T23462	31863-31869	NN	denotes	allele
T23466	31870-31871	-LRB-	denotes	(
T23467	31871-31874	JJ	denotes	top
T23468	31874-31875	-RRB-	denotes	)
T23469	31875-31877	,	denotes	,
T23470	31877-31880	DT	denotes	the
T23472	31881-31890	NN	denotes	targeting
T23471	31891-31897	NN	denotes	vector
T23473	31898-31899	-LRB-	denotes	(
T23474	31899-31905	JJ	denotes	middle
T23475	31905-31906	-RRB-	denotes	)
T23476	31906-31908	,	denotes	,
T23477	31908-31911	CC	denotes	and
T23478	31912-31915	DT	denotes	the
T23480	31916-31922	NN	denotes	mutant
T23479	31923-31929	NN	denotes	allele
T23481	31930-31931	-LRB-	denotes	(
T23482	31931-31937	JJ	denotes	bottom
T23483	31937-31938	-RRB-	denotes	)
T23484	31939-31942	VBP	denotes	are
T23459	31943-31951	VBN	denotes	depicted
T23485	31951-31952	.	denotes	.
T23486	31952-32298	sentence	denotes	In the mutant allele, a PGK-Neo cassette replaces a 0.8-kb genomic fragment containing exon 7 (grey rectangle) which encodes the active site asparagine residue (Asn286) In the targeting vector, the PGK-Neo cassette is flanked by 2.7-kb of Capn2 homologous sequence in the upstream (short) arm and 7.9-kb of homology in the downstream (long) arm.
T23487	31953-31955	IN	denotes	In
T23489	31956-31959	DT	denotes	the
T23491	31960-31966	NN	denotes	mutant
T23490	31967-31973	NN	denotes	allele
T23492	31973-31975	,	denotes	,
T23493	31975-31976	DT	denotes	a
T23495	31977-31980	NN	denotes	PGK
T23497	31980-31981	HYPH	denotes	-
T23496	31981-31984	NN	denotes	Neo
T23494	31985-31993	NN	denotes	cassette
T23488	31994-32002	VBZ	denotes	replaces
T23499	32003-32004	DT	denotes	a
T23501	32005-32008	CD	denotes	0.8
T23503	32008-32009	HYPH	denotes	-
T23502	32009-32011	NN	denotes	kb
T23504	32012-32019	JJ	denotes	genomic
T23500	32020-32028	NN	denotes	fragment
T23505	32029-32039	VBG	denotes	containing
T23506	32040-32044	NN	denotes	exon
T23507	32045-32046	CD	denotes	7
T23508	32047-32048	-LRB-	denotes	(
T23510	32048-32052	JJ	denotes	grey
T23509	32053-32062	NN	denotes	rectangle
T23511	32062-32063	-RRB-	denotes	)
T23512	32064-32069	WDT	denotes	which
T23513	32070-32077	VBZ	denotes	encodes
T23514	32078-32081	DT	denotes	the
T23516	32082-32088	JJ	denotes	active
T23517	32089-32093	NN	denotes	site
T23518	32094-32104	NN	denotes	asparagine
T23515	32105-32112	NN	denotes	residue
T23519	32113-32114	-LRB-	denotes	(
T23520	32114-32120	NN	denotes	Asn286
T23521	32120-32121	-RRB-	denotes	)
T23522	32122-32124	IN	denotes	In
T23523	32125-32128	DT	denotes	the
T23525	32129-32138	NN	denotes	targeting
T23524	32139-32145	NN	denotes	vector
T23526	32145-32147	,	denotes	,
T23527	32147-32150	DT	denotes	the
T23529	32151-32154	NN	denotes	PGK
T23531	32154-32155	HYPH	denotes	-
T23530	32155-32158	NN	denotes	Neo
T23528	32159-32167	NN	denotes	cassette
T23532	32168-32170	VBZ	denotes	is
T23498	32171-32178	VBN	denotes	flanked
T23533	32179-32181	IN	denotes	by
T23534	32182-32185	CD	denotes	2.7
T23536	32185-32186	HYPH	denotes	-
T23535	32186-32188	NN	denotes	kb
T23537	32189-32191	IN	denotes	of
T23538	32192-32197	NN	denotes	Capn2
T23540	32198-32208	JJ	denotes	homologous
T23539	32209-32217	NN	denotes	sequence
T23541	32218-32220	IN	denotes	in
T23542	32221-32224	DT	denotes	the
T23544	32225-32233	JJ	denotes	upstream
T23545	32234-32235	-LRB-	denotes	(
T23546	32235-32240	JJ	denotes	short
T23547	32240-32241	-RRB-	denotes	)
T23543	32242-32245	NN	denotes	arm
T23548	32246-32249	CC	denotes	and
T23549	32250-32253	CD	denotes	7.9
T23551	32253-32254	HYPH	denotes	-
T23550	32254-32256	NN	denotes	kb
T23552	32257-32259	IN	denotes	of
T23553	32260-32268	NN	denotes	homology
T23554	32269-32271	IN	denotes	in
T23555	32272-32275	DT	denotes	the
T23557	32276-32286	JJ	denotes	downstream
T23558	32287-32288	-LRB-	denotes	(
T23559	32288-32292	JJ	denotes	long
T23560	32292-32293	-RRB-	denotes	)
T23556	32294-32297	NN	denotes	arm
T23561	32297-32298	.	denotes	.
T23562	32298-32461	sentence	denotes	A probe located immediately outside of the short arm detects a 3.5-kb BamHI fragment from the wild-type allele and a 5.3-kb BamHI fragment from the mutant allele.
T23563	32299-32300	DT	denotes	A
T23564	32301-32306	NN	denotes	probe
T23566	32307-32314	VBN	denotes	located
T23567	32315-32326	RB	denotes	immediately
T23568	32327-32334	JJ	denotes	outside
T23569	32335-32337	IN	denotes	of
T23570	32338-32341	DT	denotes	the
T23572	32342-32347	JJ	denotes	short
T23571	32348-32351	NN	denotes	arm
T23565	32352-32359	VBZ	denotes	detects
T23573	32360-32361	DT	denotes	a
T23575	32362-32365	CD	denotes	3.5
T23577	32365-32366	HYPH	denotes	-
T23576	32366-32368	NN	denotes	kb
T23578	32369-32374	NN	denotes	BamHI
T23574	32375-32383	NN	denotes	fragment
T23579	32384-32388	IN	denotes	from
T23580	32389-32392	DT	denotes	the
T23582	32393-32397	JJ	denotes	wild
T23584	32397-32398	HYPH	denotes	-
T23583	32398-32402	NN	denotes	type
T23581	32403-32409	NN	denotes	allele
T23585	32410-32413	CC	denotes	and
T23586	32414-32415	DT	denotes	a
T23588	32416-32419	CD	denotes	5.3
T23590	32419-32420	HYPH	denotes	-
T23589	32420-32422	NN	denotes	kb
T23591	32423-32428	NN	denotes	BamHI
T23587	32429-32437	NN	denotes	fragment
T23592	32438-32442	IN	denotes	from
T23593	32443-32446	DT	denotes	the
T23595	32447-32453	NN	denotes	mutant
T23594	32454-32460	NN	denotes	allele
T23596	32460-32461	.	denotes	.
T23597	32461-32578	sentence	denotes	Exons are depicted as open vertical rectangles except for exon 7 which is represented by a solid vertical rectangle.
T23598	32462-32467	NNS	denotes	Exons
T23600	32468-32471	VBP	denotes	are
T23599	32472-32480	VBN	denotes	depicted
T23601	32481-32483	IN	denotes	as
T23602	32484-32488	JJ	denotes	open
T23604	32489-32497	JJ	denotes	vertical
T23603	32498-32508	NNS	denotes	rectangles
T23605	32509-32515	IN	denotes	except
T23606	32516-32519	IN	denotes	for
T23607	32520-32524	NN	denotes	exon
T23608	32525-32526	CD	denotes	7
T23609	32527-32532	WDT	denotes	which
T23611	32533-32535	VBZ	denotes	is
T23610	32536-32547	VBN	denotes	represented
T23612	32548-32550	IN	denotes	by
T23613	32551-32552	DT	denotes	a
T23615	32553-32558	JJ	denotes	solid
T23616	32559-32567	JJ	denotes	vertical
T23614	32568-32577	NN	denotes	rectangle
T23617	32577-32578	.	denotes	.
T23618	32578-32752	sentence	denotes	The probe used in most Southern blot analyses is shown as a solid, horizontal rectangle, while triangles mark the positions of PCR primers also used for genotyping purposes.
T23619	32579-32582	DT	denotes	The
T23620	32583-32588	NN	denotes	probe
T23622	32589-32593	VBN	denotes	used
T23623	32594-32596	IN	denotes	in
T23624	32597-32601	JJS	denotes	most
T23626	32602-32610	NNP	denotes	Southern
T23627	32611-32615	NN	denotes	blot
T23625	32616-32624	NNS	denotes	analyses
T23628	32625-32627	VBZ	denotes	is
T23621	32628-32633	VBN	denotes	shown
T23629	32634-32636	IN	denotes	as
T23630	32637-32638	DT	denotes	a
T23632	32639-32644	JJ	denotes	solid
T23633	32644-32646	,	denotes	,
T23634	32646-32656	JJ	denotes	horizontal
T23631	32657-32666	NN	denotes	rectangle
T23635	32666-32668	,	denotes	,
T23636	32668-32673	IN	denotes	while
T23638	32674-32683	NNS	denotes	triangles
T23637	32684-32688	VBP	denotes	mark
T23639	32689-32692	DT	denotes	the
T23640	32693-32702	NNS	denotes	positions
T23641	32703-32705	IN	denotes	of
T23642	32706-32709	NN	denotes	PCR
T23643	32710-32717	NNS	denotes	primers
T23644	32718-32722	RB	denotes	also
T23645	32723-32727	VBN	denotes	used
T23646	32728-32731	IN	denotes	for
T23647	32732-32742	NN	denotes	genotyping
T23648	32743-32751	NNS	denotes	purposes
T23649	32751-32752	.	denotes	.
T17462	32754-32756	NN	denotes	ES
T17464	32757-32761	NN	denotes	cell
T17463	32762-32769	NN	denotes	culture
T17465	32769-32771	,	denotes	,
T17466	32771-32783	NN	denotes	transfection
T17467	32784-32787	CC	denotes	and
T17468	32788-32797	NN	denotes	selection
T17469	32798-32800	IN	denotes	of
T17470	32801-32809	VBN	denotes	targeted
T17471	32810-32816	NNS	denotes	clones
T17472	32816-33235	sentence	denotes	Mouse R1 ES cells [49] were maintained on gelatin-coated plates with feeder layers of mouse embryonic fibroblasts at 37°C under 5% CO2 in ES cell medium (DMEM [high glucose] supplemented with 15% fetal bovine serum, 0.1 mM non-essential amino acids, 2 mM glutamine, 1 mM sodium pyruvate, 0.1 mM 2-mercaptoethanol, 100 U/ml penicillin, 100 μg/ml streptomycin, 25 μg/ml amphotericin, and 1,000 U/ml of ESGro [Chemicon]).
T17473	32817-32822	NN	denotes	Mouse
T17475	32823-32825	NN	denotes	R1
T17476	32826-32828	NN	denotes	ES
T17474	32829-32834	NNS	denotes	cells
T17478	32835-32836	-LRB-	denotes	[
T17479	32836-32838	CD	denotes	49
T17480	32838-32839	-RRB-	denotes	]
T17481	32840-32844	VBD	denotes	were
T17477	32845-32855	VBN	denotes	maintained
T17482	32856-32858	IN	denotes	on
T17483	32859-32866	NN	denotes	gelatin
T17485	32866-32867	HYPH	denotes	-
T17484	32867-32873	VBN	denotes	coated
T17486	32874-32880	NNS	denotes	plates
T17487	32881-32885	IN	denotes	with
T17488	32886-32892	NN	denotes	feeder
T17489	32893-32899	NNS	denotes	layers
T17490	32900-32902	IN	denotes	of
T17491	32903-32908	NN	denotes	mouse
T17493	32909-32918	JJ	denotes	embryonic
T17492	32919-32930	NNS	denotes	fibroblasts
T17494	32931-32933	IN	denotes	at
T17495	32934-32936	CD	denotes	37
T17496	32936-32938	NN	denotes	°C
T17497	32939-32944	IN	denotes	under
T17498	32945-32946	CD	denotes	5
T17499	32946-32947	NN	denotes	%
T17500	32948-32951	NN	denotes	CO2
T17501	32952-32954	IN	denotes	in
T17502	32955-32957	NN	denotes	ES
T17504	32958-32962	NN	denotes	cell
T17503	32963-32969	NN	denotes	medium
T17505	32970-32971	-LRB-	denotes	(
T17506	32971-32975	NN	denotes	DMEM
T17507	32976-32977	-LRB-	denotes	[
T17509	32977-32981	JJ	denotes	high
T17508	32982-32989	NN	denotes	glucose
T17510	32989-32990	-RRB-	denotes	]
T17511	32991-33003	VBN	denotes	supplemented
T17512	33004-33008	IN	denotes	with
T17513	33009-33011	CD	denotes	15
T17514	33011-33012	NN	denotes	%
T17516	33013-33018	JJ	denotes	fetal
T17517	33019-33025	JJ	denotes	bovine
T17515	33026-33031	NN	denotes	serum
T17518	33031-33033	,	denotes	,
T17519	33033-33036	CD	denotes	0.1
T17520	33037-33039	NN	denotes	mM
T17522	33040-33053	JJ	denotes	non-essential
T17523	33054-33059	NN	denotes	amino
T17521	33060-33065	NNS	denotes	acids
T17524	33065-33067	,	denotes	,
T17525	33067-33068	CD	denotes	2
T17526	33069-33071	NN	denotes	mM
T17527	33072-33081	NN	denotes	glutamine
T17528	33081-33083	,	denotes	,
T17529	33083-33084	CD	denotes	1
T17530	33085-33087	NN	denotes	mM
T17532	33088-33094	NN	denotes	sodium
T17531	33095-33103	NN	denotes	pyruvate
T17533	33103-33105	,	denotes	,
T17534	33105-33108	CD	denotes	0.1
T17535	33109-33111	NN	denotes	mM
T17537	33112-33113	CD	denotes	2
T17538	33113-33114	HYPH	denotes	-
T17536	33114-33129	NN	denotes	mercaptoethanol
T17539	33129-33131	,	denotes	,
T17540	33131-33134	CD	denotes	100
T17541	33135-33136	NN	denotes	U
T17543	33136-33137	SYM	denotes	/
T17544	33137-33139	NN	denotes	ml
T17542	33140-33150	NN	denotes	penicillin
T17545	33150-33152	,	denotes	,
T17546	33152-33155	CD	denotes	100
T17547	33156-33158	NN	denotes	μg
T17549	33158-33159	SYM	denotes	/
T17550	33159-33161	NN	denotes	ml
T17548	33162-33174	NN	denotes	streptomycin
T17551	33174-33176	,	denotes	,
T17552	33176-33178	CD	denotes	25
T17553	33179-33181	NN	denotes	μg
T17555	33181-33182	SYM	denotes	/
T17556	33182-33184	NN	denotes	ml
T17554	33185-33197	NN	denotes	amphotericin
T17557	33197-33199	,	denotes	,
T17558	33199-33202	CC	denotes	and
T17559	33203-33208	CD	denotes	1,000
T17560	33209-33210	NN	denotes	U
T17561	33210-33211	SYM	denotes	/
T17562	33211-33213	NN	denotes	ml
T17563	33214-33216	IN	denotes	of
T17564	33217-33222	NN	denotes	ESGro
T17565	33223-33224	-LRB-	denotes	[
T17566	33224-33232	NNP	denotes	Chemicon
T17567	33232-33233	-RRB-	denotes	]
T17568	33233-33234	-RRB-	denotes	)
T17569	33234-33235	.	denotes	.
T17570	33235-33352	sentence	denotes	Fetal bovine serum from HyClone Laboratories Inc (Logan, Utah) was tested for its ability to support ES cell growth.
T17571	33236-33241	JJ	denotes	Fetal
T17573	33242-33248	JJ	denotes	bovine
T17572	33249-33254	NN	denotes	serum
T17575	33255-33259	IN	denotes	from
T17576	33260-33267	NNP	denotes	HyClone
T17578	33268-33280	NNP	denotes	Laboratories
T17577	33281-33284	NNP	denotes	Inc
T17579	33285-33286	-LRB-	denotes	(
T17580	33286-33291	NNP	denotes	Logan
T17581	33291-33293	,	denotes	,
T17582	33293-33297	NNP	denotes	Utah
T17583	33297-33298	-RRB-	denotes	)
T17584	33299-33302	VBD	denotes	was
T17574	33303-33309	VBN	denotes	tested
T17585	33310-33313	IN	denotes	for
T17586	33314-33317	PRP$	denotes	its
T17587	33318-33325	NN	denotes	ability
T17588	33326-33328	TO	denotes	to
T17589	33329-33336	VB	denotes	support
T17590	33337-33339	NN	denotes	ES
T17591	33340-33344	NN	denotes	cell
T17592	33345-33351	NN	denotes	growth
T17593	33351-33352	.	denotes	.
T17594	33352-33411	sentence	denotes	Gelatin was from Sigma-Aldrich Canada (Oakville, Ontario).
T17595	33353-33360	NN	denotes	Gelatin
T17596	33361-33364	VBD	denotes	was
T17597	33365-33369	IN	denotes	from
T17598	33370-33375	NNP	denotes	Sigma
T17600	33375-33376	HYPH	denotes	-
T17599	33376-33383	NNP	denotes	Aldrich
T17601	33384-33390	NNP	denotes	Canada
T17602	33391-33392	-LRB-	denotes	(
T17603	33392-33400	NNP	denotes	Oakville
T17604	33400-33402	,	denotes	,
T17605	33402-33409	NNP	denotes	Ontario
T17606	33409-33410	-RRB-	denotes	)
T17607	33410-33411	.	denotes	.
T17608	33411-33494	sentence	denotes	Unless otherwise specified, all other tissue culture reagents were from Gibco-BRL.
T17609	33412-33418	IN	denotes	Unless
T17611	33419-33428	RB	denotes	otherwise
T17610	33429-33438	VBN	denotes	specified
T17613	33438-33440	,	denotes	,
T17614	33440-33443	DT	denotes	all
T17616	33444-33449	JJ	denotes	other
T17617	33450-33456	NN	denotes	tissue
T17618	33457-33464	NN	denotes	culture
T17615	33465-33473	NNS	denotes	reagents
T17612	33474-33478	VBD	denotes	were
T17619	33479-33483	IN	denotes	from
T17620	33484-33489	NNP	denotes	Gibco
T17622	33489-33490	HYPH	denotes	-
T17621	33490-33493	NNP	denotes	BRL
T17623	33493-33494	.	denotes	.
T17624	33494-33588	sentence	denotes	The targeting construct was linearized by NotI digestion and electroporated into R1 ES cells.
T17625	33495-33498	DT	denotes	The
T17627	33499-33508	NN	denotes	targeting
T17626	33509-33518	NN	denotes	construct
T17629	33519-33522	VBD	denotes	was
T17628	33523-33533	VBN	denotes	linearized
T17630	33534-33536	IN	denotes	by
T17631	33537-33541	NN	denotes	NotI
T17632	33542-33551	NN	denotes	digestion
T17633	33552-33555	CC	denotes	and
T17634	33556-33570	VBN	denotes	electroporated
T17635	33571-33575	IN	denotes	into
T17636	33576-33578	NN	denotes	R1
T17638	33579-33581	NN	denotes	ES
T17637	33582-33587	NNS	denotes	cells
T17639	33587-33588	.	denotes	.
T17640	33588-33789	sentence	denotes	Cells were plated without feeder layers on gelatin-coated plates and transformed clones were selected in the presence of 200 μg/ml G418 (Gibco- BRL) and 2 μM ganciclovir (Syntex, Inc.) for eight days.
T17641	33589-33594	NNS	denotes	Cells
T17643	33595-33599	VBD	denotes	were
T17642	33600-33606	VBN	denotes	plated
T17644	33607-33614	IN	denotes	without
T17645	33615-33621	NN	denotes	feeder
T17646	33622-33628	NNS	denotes	layers
T17647	33629-33631	IN	denotes	on
T17648	33632-33639	NN	denotes	gelatin
T17650	33639-33640	HYPH	denotes	-
T17649	33640-33646	VBN	denotes	coated
T17651	33647-33653	NNS	denotes	plates
T17652	33654-33657	CC	denotes	and
T17653	33658-33669	VBN	denotes	transformed
T17654	33670-33676	NNS	denotes	clones
T17656	33677-33681	VBD	denotes	were
T17655	33682-33690	VBN	denotes	selected
T17657	33691-33693	IN	denotes	in
T17658	33694-33697	DT	denotes	the
T17659	33698-33706	NN	denotes	presence
T17660	33707-33709	IN	denotes	of
T17661	33710-33713	CD	denotes	200
T17662	33714-33716	NN	denotes	μg
T17664	33716-33717	SYM	denotes	/
T17665	33717-33719	NN	denotes	ml
T17663	33720-33724	NN	denotes	G418
T17666	33725-33726	-LRB-	denotes	(
T17668	33726-33731	NNP	denotes	Gibco
T17669	33731-33732	HYPH	denotes	-
T17667	33733-33736	NNP	denotes	BRL
T17670	33736-33737	-RRB-	denotes	)
T17671	33738-33741	CC	denotes	and
T17672	33742-33743	CD	denotes	2
T17673	33744-33746	NN	denotes	μM
T17674	33747-33758	NN	denotes	ganciclovir
T17675	33759-33760	-LRB-	denotes	(
T17676	33760-33766	NNP	denotes	Syntex
T17677	33766-33768	,	denotes	,
T17678	33768-33772	NNP	denotes	Inc.
T17679	33772-33773	-RRB-	denotes	)
T17680	33774-33777	IN	denotes	for
T17681	33778-33783	CD	denotes	eight
T17682	33784-33788	NNS	denotes	days
T17683	33788-33789	.	denotes	.
T17684	33789-33924	sentence	denotes	Drug-resistant clones were picked, expanded on gelatin-coated plates, and genotyped by Southern blotting and PCR analysis (see below).
T17685	33790-33794	NN	denotes	Drug
T17687	33794-33795	HYPH	denotes	-
T17686	33795-33804	JJ	denotes	resistant
T17688	33805-33811	NNS	denotes	clones
T17690	33812-33816	VBD	denotes	were
T17689	33817-33823	VBN	denotes	picked
T17691	33823-33825	,	denotes	,
T17692	33825-33833	VBN	denotes	expanded
T17693	33834-33836	IN	denotes	on
T17694	33837-33844	NN	denotes	gelatin
T17696	33844-33845	HYPH	denotes	-
T17695	33845-33851	VBN	denotes	coated
T17697	33852-33858	NNS	denotes	plates
T17698	33858-33860	,	denotes	,
T17699	33860-33863	CC	denotes	and
T17700	33864-33873	VBN	denotes	genotyped
T17701	33874-33876	IN	denotes	by
T17702	33877-33885	NNP	denotes	Southern
T17703	33886-33894	NN	denotes	blotting
T17704	33895-33898	CC	denotes	and
T17705	33899-33902	NN	denotes	PCR
T17706	33903-33911	NN	denotes	analysis
T17707	33912-33913	-LRB-	denotes	(
T17708	33913-33916	VB	denotes	see
T17709	33917-33922	RB	denotes	below
T17710	33922-33923	-RRB-	denotes	)
T17711	33923-33924	.	denotes	.
T17988	33926-33936	NN	denotes	Generation
T17989	33937-33939	IN	denotes	of
T17990	33940-33948	VBN	denotes	targeted
T17991	33949-33953	NNS	denotes	mice
T17992	33953-34075	sentence	denotes	Capn2+/- ES cells were aggregated overnight with 8-cell embryos recovered from CD1 matings, as previously described [31].
T17993	33954-33959	NN	denotes	Capn2
T17995	33959-33960	SYM	denotes	+
T17996	33960-33961	HYPH	denotes	/
T17997	33961-33962	SYM	denotes	-
T17998	33963-33965	NN	denotes	ES
T17994	33966-33971	NNS	denotes	cells
T18000	33972-33976	VBD	denotes	were
T17999	33977-33987	VBN	denotes	aggregated
T18001	33988-33997	RB	denotes	overnight
T18002	33998-34002	IN	denotes	with
T18003	34003-34004	CD	denotes	8
T18005	34004-34005	HYPH	denotes	-
T18004	34005-34009	NN	denotes	cell
T18006	34010-34017	NNS	denotes	embryos
T18007	34018-34027	VBN	denotes	recovered
T18008	34028-34032	IN	denotes	from
T18009	34033-34036	NN	denotes	CD1
T18010	34037-34044	NNS	denotes	matings
T18011	34044-34046	,	denotes	,
T18012	34046-34048	IN	denotes	as
T18014	34049-34059	RB	denotes	previously
T18013	34060-34069	VBN	denotes	described
T18015	34070-34071	-LRB-	denotes	[
T18016	34071-34073	CD	denotes	31
T18017	34073-34074	-RRB-	denotes	]
T18018	34074-34075	.	denotes	.
T18019	34075-34152	sentence	denotes	On the next day, blastocysts were transferred to pseudopregnant CD1 females.
T18020	34076-34078	IN	denotes	On
T18022	34079-34082	DT	denotes	the
T18024	34083-34087	JJ	denotes	next
T18023	34088-34091	NN	denotes	day
T18025	34091-34093	,	denotes	,
T18026	34093-34104	NNS	denotes	blastocysts
T18027	34105-34109	VBD	denotes	were
T18021	34110-34121	VBN	denotes	transferred
T18028	34122-34124	IN	denotes	to
T18029	34125-34139	JJ	denotes	pseudopregnant
T18031	34140-34143	NN	denotes	CD1
T18030	34144-34151	NNS	denotes	females
T18032	34151-34152	.	denotes	.
T18033	34152-34273	sentence	denotes	Chimeric animals were identifiable at birth by black eye pigmentation and subsequently by patches of agouti coat colour.
T18034	34153-34161	JJ	denotes	Chimeric
T18035	34162-34169	NNS	denotes	animals
T18036	34170-34174	VBD	denotes	were
T18037	34175-34187	JJ	denotes	identifiable
T18038	34188-34190	IN	denotes	at
T18039	34191-34196	NN	denotes	birth
T18040	34197-34199	IN	denotes	by
T18041	34200-34205	JJ	denotes	black
T18043	34206-34209	NN	denotes	eye
T18042	34210-34222	NN	denotes	pigmentation
T18044	34223-34226	CC	denotes	and
T18045	34227-34239	RB	denotes	subsequently
T18046	34240-34242	IN	denotes	by
T18047	34243-34250	NNS	denotes	patches
T18048	34251-34253	IN	denotes	of
T18049	34254-34260	NN	denotes	agouti
T18051	34261-34265	NN	denotes	coat
T18050	34266-34272	NN	denotes	colour
T18052	34272-34273	.	denotes	.
T18053	34273-34373	sentence	denotes	Chimeric males were bred with CD1 females to identify those males capable of germline transmission.
T18054	34274-34282	JJ	denotes	Chimeric
T18055	34283-34288	NNS	denotes	males
T18057	34289-34293	VBD	denotes	were
T18056	34294-34298	VBN	denotes	bred
T18058	34299-34303	IN	denotes	with
T18059	34304-34307	NN	denotes	CD1
T18060	34308-34315	NNS	denotes	females
T18061	34316-34318	TO	denotes	to
T18062	34319-34327	VB	denotes	identify
T18063	34328-34333	DT	denotes	those
T18064	34334-34339	NNS	denotes	males
T18065	34340-34347	JJ	denotes	capable
T18066	34348-34350	IN	denotes	of
T18067	34351-34359	NN	denotes	germline
T18068	34360-34372	NN	denotes	transmission
T18069	34372-34373	.	denotes	.
T18070	34373-34473	sentence	denotes	These were then bred with 129SvJ females to establish the mutation in an inbred genetic background.
T18071	34374-34379	DT	denotes	These
T18073	34380-34384	VBD	denotes	were
T18074	34385-34389	RB	denotes	then
T18072	34390-34394	VBN	denotes	bred
T18075	34395-34399	IN	denotes	with
T18076	34400-34406	NN	denotes	129SvJ
T18077	34407-34414	NNS	denotes	females
T18078	34415-34417	TO	denotes	to
T18079	34418-34427	VB	denotes	establish
T18080	34428-34431	DT	denotes	the
T18081	34432-34440	NN	denotes	mutation
T18082	34441-34443	IN	denotes	in
T18083	34444-34446	DT	denotes	an
T18085	34447-34453	JJ	denotes	inbred
T18086	34454-34461	JJ	denotes	genetic
T18084	34462-34472	NN	denotes	background
T18087	34472-34473	.	denotes	.
T18088	34473-34619	sentence	denotes	Mouse protocols were approved by the Queen's University Animal Care Committee according to the guidelines of the Canadian Council on Animal Care.
T18089	34474-34479	NN	denotes	Mouse
T18090	34480-34489	NNS	denotes	protocols
T18092	34490-34494	VBD	denotes	were
T18091	34495-34503	VBN	denotes	approved
T18093	34504-34506	IN	denotes	by
T18094	34507-34510	DT	denotes	the
T18095	34511-34516	NNP	denotes	Queen
T18097	34516-34518	POS	denotes	's
T18096	34519-34529	NNP	denotes	University
T18099	34530-34536	NNP	denotes	Animal
T18100	34537-34541	NNP	denotes	Care
T18098	34542-34551	NNP	denotes	Committee
T18101	34552-34561	VBG	denotes	according
T18102	34562-34564	IN	denotes	to
T18103	34565-34568	DT	denotes	the
T18104	34569-34579	NNS	denotes	guidelines
T18105	34580-34582	IN	denotes	of
T18106	34583-34586	DT	denotes	the
T18108	34587-34595	NNP	denotes	Canadian
T18107	34596-34603	NNP	denotes	Council
T18109	34604-34606	IN	denotes	on
T18110	34607-34613	NNP	denotes	Animal
T18111	34614-34618	NNP	denotes	Care
T18112	34618-34619	.	denotes	.
T19530	34621-34631	NN	denotes	Genotyping
T19531	34632-34639	NNS	denotes	methods
T19532	34639-34750	sentence	denotes	Several Southern blot and PCR strategies were exploited in order to determine the genotype of the Capn2 locus.
T19533	34640-34647	JJ	denotes	Several
T19535	34648-34656	NNP	denotes	Southern
T19534	34657-34661	NN	denotes	blot
T19537	34662-34665	CC	denotes	and
T19538	34666-34669	NN	denotes	PCR
T19539	34670-34680	NNS	denotes	strategies
T19540	34681-34685	VBD	denotes	were
T19536	34686-34695	VBN	denotes	exploited
T19541	34696-34698	IN	denotes	in
T19542	34699-34704	NN	denotes	order
T19543	34705-34707	TO	denotes	to
T19544	34708-34717	VB	denotes	determine
T19545	34718-34721	DT	denotes	the
T19546	34722-34730	NN	denotes	genotype
T19547	34731-34733	IN	denotes	of
T19548	34734-34737	DT	denotes	the
T19550	34738-34743	NN	denotes	Capn2
T19549	34744-34749	NN	denotes	locus
T19551	34749-34750	.	denotes	.
T19552	34750-34844	sentence	denotes	Southern blotting was carried out using the digoxigenin (DIG) non-radioactive system (Roche).
T19553	34751-34759	NNP	denotes	Southern
T19554	34760-34768	NN	denotes	blotting
T19556	34769-34772	VBD	denotes	was
T19555	34773-34780	VBN	denotes	carried
T19557	34781-34784	RP	denotes	out
T19558	34785-34790	VBG	denotes	using
T19559	34791-34794	DT	denotes	the
T19561	34795-34806	NN	denotes	digoxigenin
T19562	34807-34808	-LRB-	denotes	(
T19563	34808-34811	NN	denotes	DIG
T19564	34811-34812	-RRB-	denotes	)
T19565	34813-34828	JJ	denotes	non-radioactive
T19560	34829-34835	NN	denotes	system
T19566	34836-34837	-LRB-	denotes	(
T19567	34837-34842	NNP	denotes	Roche
T19568	34842-34843	-RRB-	denotes	)
T19569	34843-34844	.	denotes	.
T19570	34844-35066	sentence	denotes	In most cases, membranes were blotted with BamHI-digested genomic DNA and hybridized with a DIG-labeled 823 bp exon 4-containing BamHI-HindIII fragment located immediately upstream of the short arm of homology (Figure 1).
T19571	34845-34847	IN	denotes	In
T19573	34848-34852	JJS	denotes	most
T19574	34853-34858	NNS	denotes	cases
T19575	34858-34860	,	denotes	,
T19576	34860-34869	NNS	denotes	membranes
T19577	34870-34874	VBD	denotes	were
T19572	34875-34882	VBN	denotes	blotted
T19578	34883-34887	IN	denotes	with
T19579	34888-34893	NN	denotes	BamHI
T19581	34893-34894	HYPH	denotes	-
T19580	34894-34902	VBN	denotes	digested
T19583	34903-34910	JJ	denotes	genomic
T19582	34911-34914	NN	denotes	DNA
T19584	34915-34918	CC	denotes	and
T19585	34919-34929	VBD	denotes	hybridized
T19586	34930-34934	IN	denotes	with
T19587	34935-34936	DT	denotes	a
T19589	34937-34940	NN	denotes	DIG
T19591	34940-34941	HYPH	denotes	-
T19590	34941-34948	VBN	denotes	labeled
T19592	34949-34952	CD	denotes	823
T19593	34953-34955	NN	denotes	bp
T19594	34956-34960	NN	denotes	exon
T19596	34961-34962	CD	denotes	4
T19597	34962-34963	HYPH	denotes	-
T19595	34963-34973	VBG	denotes	containing
T19598	34974-34979	NN	denotes	BamHI
T19600	34979-34980	HYPH	denotes	-
T19599	34980-34987	NN	denotes	HindIII
T19588	34988-34996	NN	denotes	fragment
T19601	34997-35004	VBN	denotes	located
T19602	35005-35016	RB	denotes	immediately
T19603	35017-35025	RB	denotes	upstream
T19604	35026-35028	IN	denotes	of
T19605	35029-35032	DT	denotes	the
T19607	35033-35038	JJ	denotes	short
T19606	35039-35042	NN	denotes	arm
T19608	35043-35045	IN	denotes	of
T19609	35046-35054	NN	denotes	homology
T19610	35055-35056	-LRB-	denotes	(
T19611	35056-35062	NN	denotes	Figure
T19612	35063-35064	CD	denotes	1
T19613	35064-35065	-RRB-	denotes	)
T19614	35065-35066	.	denotes	.
T19615	35066-35224	sentence	denotes	A 681 bp PstI-XbaI fragment from the PGK-Neo cassette was also used to probe Southern blots in order to verify a single integration event in targeted clones.
T19616	35067-35068	DT	denotes	A
T19618	35069-35072	CD	denotes	681
T19619	35073-35075	NN	denotes	bp
T19620	35076-35080	NN	denotes	PstI
T19622	35080-35081	HYPH	denotes	-
T19621	35081-35085	NN	denotes	XbaI
T19617	35086-35094	NN	denotes	fragment
T19624	35095-35099	IN	denotes	from
T19625	35100-35103	DT	denotes	the
T19627	35104-35107	NN	denotes	PGK
T19629	35107-35108	HYPH	denotes	-
T19628	35108-35111	NN	denotes	Neo
T19626	35112-35120	NN	denotes	cassette
T19630	35121-35124	VBD	denotes	was
T19631	35125-35129	RB	denotes	also
T19623	35130-35134	VBN	denotes	used
T19632	35135-35137	TO	denotes	to
T19633	35138-35143	VB	denotes	probe
T19634	35144-35152	NNP	denotes	Southern
T19635	35153-35158	NNS	denotes	blots
T19636	35159-35161	IN	denotes	in
T19637	35162-35167	NN	denotes	order
T19638	35168-35170	TO	denotes	to
T19639	35171-35177	VB	denotes	verify
T19640	35178-35179	DT	denotes	a
T19642	35180-35186	JJ	denotes	single
T19643	35187-35198	NN	denotes	integration
T19641	35199-35204	NN	denotes	event
T19644	35205-35207	IN	denotes	in
T19645	35208-35216	VBN	denotes	targeted
T19646	35217-35223	NNS	denotes	clones
T19647	35223-35224	.	denotes	.
T19648	35224-35288	sentence	denotes	Genotyping was also carried out by PCR analysis of genomic DNA.
T19649	35225-35235	NN	denotes	Genotyping
T19651	35236-35239	VBD	denotes	was
T19652	35240-35244	RB	denotes	also
T19650	35245-35252	VBN	denotes	carried
T19653	35253-35256	RP	denotes	out
T19654	35257-35259	IN	denotes	by
T19655	35260-35263	NN	denotes	PCR
T19656	35264-35272	NN	denotes	analysis
T19657	35273-35275	IN	denotes	of
T19658	35276-35283	JJ	denotes	genomic
T19659	35284-35287	NN	denotes	DNA
T19660	35287-35288	.	denotes	.
T19661	35288-35356	sentence	denotes	The sequences of all oligonucleotide primers are listed in Table 1.
T19662	35289-35292	DT	denotes	The
T19663	35293-35302	NNS	denotes	sequences
T19665	35303-35305	IN	denotes	of
T19666	35306-35309	DT	denotes	all
T19668	35310-35325	NN	denotes	oligonucleotide
T19667	35326-35333	NNS	denotes	primers
T19669	35334-35337	VBP	denotes	are
T19664	35338-35344	VBN	denotes	listed
T19670	35345-35347	IN	denotes	in
T19671	35348-35353	NN	denotes	Table
T19672	35354-35355	CD	denotes	1
T19673	35355-35356	.	denotes	.
T19674	35356-35491	sentence	denotes	A single-step PCR strategy was sufficient for genotyping ES cells or biopsies from post-implantation embryos and weanlings (Figure 1).
T19675	35357-35358	DT	denotes	A
T19677	35359-35365	JJ	denotes	single
T19679	35365-35366	HYPH	denotes	-
T19678	35366-35370	NN	denotes	step
T19680	35371-35374	NN	denotes	PCR
T19676	35375-35383	NN	denotes	strategy
T19681	35384-35387	VBD	denotes	was
T19682	35388-35398	JJ	denotes	sufficient
T19683	35399-35402	IN	denotes	for
T19684	35403-35413	VBG	denotes	genotyping
T19685	35414-35416	NN	denotes	ES
T19686	35417-35422	NNS	denotes	cells
T19687	35423-35425	CC	denotes	or
T19688	35426-35434	NNS	denotes	biopsies
T19689	35435-35439	IN	denotes	from
T19690	35440-35457	JJ	denotes	post-implantation
T19691	35458-35465	NNS	denotes	embryos
T19692	35466-35469	CC	denotes	and
T19693	35470-35479	NNS	denotes	weanlings
T19694	35480-35481	-LRB-	denotes	(
T19695	35481-35487	NN	denotes	Figure
T19696	35488-35489	CD	denotes	1
T19697	35489-35490	-RRB-	denotes	)
T19698	35490-35491	.	denotes	.
T19699	35491-35815	sentence	denotes	A 2,748 bp segment of the wild-type allele and a 2,711 bp segment of the mutant allele were amplified in separate reactions using a common (intron 4) sense primer, located outside the short arm of homology, and distinct antisense primers which hybridized to either wild-type (exon 7) or mutant (PGK-Neo) sequence (Table I).
T19700	35492-35493	DT	denotes	A
T19702	35494-35499	CD	denotes	2,748
T19703	35500-35502	NN	denotes	bp
T19701	35503-35510	NN	denotes	segment
T19705	35511-35513	IN	denotes	of
T19706	35514-35517	DT	denotes	the
T19708	35518-35522	JJ	denotes	wild
T19710	35522-35523	HYPH	denotes	-
T19709	35523-35527	NN	denotes	type
T19707	35528-35534	NN	denotes	allele
T19711	35535-35538	CC	denotes	and
T19712	35539-35540	DT	denotes	a
T19714	35541-35546	CD	denotes	2,711
T19715	35547-35549	NN	denotes	bp
T19713	35550-35557	NN	denotes	segment
T19716	35558-35560	IN	denotes	of
T19717	35561-35564	DT	denotes	the
T19719	35565-35571	NN	denotes	mutant
T19718	35572-35578	NN	denotes	allele
T19720	35579-35583	VBD	denotes	were
T19704	35584-35593	VBN	denotes	amplified
T19721	35594-35596	IN	denotes	in
T19722	35597-35605	JJ	denotes	separate
T19723	35606-35615	NNS	denotes	reactions
T19724	35616-35621	VBG	denotes	using
T19725	35622-35623	DT	denotes	a
T19727	35624-35630	JJ	denotes	common
T19729	35631-35632	-LRB-	denotes	(
T19730	35632-35638	NN	denotes	intron
T19731	35639-35640	CD	denotes	4
T19732	35640-35641	-RRB-	denotes	)
T19728	35642-35647	NN	denotes	sense
T19726	35648-35654	NN	denotes	primer
T19733	35654-35656	,	denotes	,
T19734	35656-35663	VBN	denotes	located
T19735	35664-35671	IN	denotes	outside
T19736	35672-35675	DT	denotes	the
T19738	35676-35681	JJ	denotes	short
T19737	35682-35685	NN	denotes	arm
T19739	35686-35688	IN	denotes	of
T19740	35689-35697	NN	denotes	homology
T19741	35697-35699	,	denotes	,
T19742	35699-35702	CC	denotes	and
T19743	35703-35711	JJ	denotes	distinct
T19745	35712-35721	NN	denotes	antisense
T19744	35722-35729	NNS	denotes	primers
T19746	35730-35735	WDT	denotes	which
T19747	35736-35746	VBD	denotes	hybridized
T19748	35747-35749	IN	denotes	to
T19749	35750-35756	CC	denotes	either
T19751	35757-35761	JJ	denotes	wild
T19752	35761-35762	HYPH	denotes	-
T19750	35762-35766	NN	denotes	type
T19754	35767-35768	-LRB-	denotes	(
T19755	35768-35772	NN	denotes	exon
T19756	35773-35774	CD	denotes	7
T19757	35774-35775	-RRB-	denotes	)
T19758	35776-35778	CC	denotes	or
T19759	35779-35785	NN	denotes	mutant
T19760	35786-35787	-LRB-	denotes	(
T19762	35787-35790	NN	denotes	PGK
T19763	35790-35791	HYPH	denotes	-
T19761	35791-35794	NN	denotes	Neo
T19764	35794-35795	-RRB-	denotes	)
T19753	35796-35804	NN	denotes	sequence
T19765	35805-35806	-LRB-	denotes	(
T19767	35806-35811	NN	denotes	Table
T19766	35812-35813	NN	denotes	I
T19768	35813-35814	-RRB-	denotes	)
T19769	35814-35815	.	denotes	.
T19770	35815-36052	sentence	denotes	The thermocycling parameters included a five minute initial denaturation step at 95°C, 30 cycles of one minute denaturation at 95°C, one minute annealing at 56°C, and one minute extension at 72°C, with a ten minute final extension step.
T19771	35816-35819	DT	denotes	The
T19773	35820-35833	VBG	denotes	thermocycling
T19772	35834-35844	NNS	denotes	parameters
T19774	35845-35853	VBD	denotes	included
T19775	35854-35855	DT	denotes	a
T19777	35856-35860	CD	denotes	five
T19778	35861-35867	NN	denotes	minute
T19779	35868-35875	JJ	denotes	initial
T19780	35876-35888	NN	denotes	denaturation
T19776	35889-35893	NN	denotes	step
T19781	35894-35896	IN	denotes	at
T19782	35897-35899	CD	denotes	95
T19783	35899-35901	NN	denotes	°C
T19784	35901-35903	,	denotes	,
T19785	35903-35905	CD	denotes	30
T19786	35906-35912	NNS	denotes	cycles
T19787	35913-35915	IN	denotes	of
T19788	35916-35919	CD	denotes	one
T19789	35920-35926	NN	denotes	minute
T19790	35927-35939	NN	denotes	denaturation
T19791	35940-35942	IN	denotes	at
T19792	35943-35945	CD	denotes	95
T19793	35945-35947	NN	denotes	°C
T19794	35947-35949	,	denotes	,
T19795	35949-35952	CD	denotes	one
T19796	35953-35959	NN	denotes	minute
T19797	35960-35969	VBG	denotes	annealing
T19798	35970-35972	IN	denotes	at
T19799	35973-35975	CD	denotes	56
T19800	35975-35977	NN	denotes	°C
T19801	35977-35979	,	denotes	,
T19802	35979-35982	CC	denotes	and
T19803	35983-35986	CD	denotes	one
T19804	35987-35993	NN	denotes	minute
T19805	35994-36003	NN	denotes	extension
T19806	36004-36006	IN	denotes	at
T19807	36007-36009	CD	denotes	72
T19808	36009-36011	NN	denotes	°C
T19809	36011-36013	,	denotes	,
T19810	36013-36017	IN	denotes	with
T19811	36018-36019	DT	denotes	a
T19813	36020-36023	CD	denotes	ten
T19814	36024-36030	NN	denotes	minute
T19815	36031-36036	JJ	denotes	final
T19816	36037-36046	NN	denotes	extension
T19812	36047-36051	NN	denotes	step
T19817	36051-36052	.	denotes	.
T19818	36052-36053	sentence	denotes
T23673	36062-36077	NN	denotes	Oligonucleotide
T23674	36078-36085	NNS	denotes	primers
T23675	36086-36090	VBN	denotes	used
T23676	36091-36093	TO	denotes	to
T23677	36094-36102	VB	denotes	genotype
T23678	36103-36106	DT	denotes	the
T23680	36107-36112	NN	denotes	Capn2
T23679	36113-36118	NN	denotes	locus
T19820	37044-37047	IN	denotes	Due
T19819	37044-37217	sentence	denotes	Due to the limited amount of genetic material available in pre-implantation embryos, a nested PCR strategy was developed to yield reliable genotyping information (Figure 4).
T19822	37048-37050	IN	denotes	to
T19823	37051-37054	DT	denotes	the
T19825	37055-37062	VBN	denotes	limited
T19824	37063-37069	NN	denotes	amount
T19826	37070-37072	IN	denotes	of
T19827	37073-37080	JJ	denotes	genetic
T19828	37081-37089	NN	denotes	material
T19829	37090-37099	JJ	denotes	available
T19830	37100-37102	IN	denotes	in
T19831	37103-37119	JJ	denotes	pre-implantation
T19832	37120-37127	NNS	denotes	embryos
T19833	37127-37129	,	denotes	,
T19834	37129-37130	DT	denotes	a
T19836	37131-37137	VBN	denotes	nested
T19837	37138-37141	NN	denotes	PCR
T19835	37142-37150	NN	denotes	strategy
T19838	37151-37154	VBD	denotes	was
T19821	37155-37164	VBN	denotes	developed
T19839	37165-37167	TO	denotes	to
T19840	37168-37173	VB	denotes	yield
T19841	37174-37182	JJ	denotes	reliable
T19843	37183-37193	NN	denotes	genotyping
T19842	37194-37205	NN	denotes	information
T19844	37206-37207	-LRB-	denotes	(
T19845	37207-37213	NN	denotes	Figure
T19846	37214-37215	CD	denotes	4
T19847	37215-37216	-RRB-	denotes	)
T19848	37216-37217	.	denotes	.
T19849	37217-37296	sentence	denotes	Whole embryos were first digested in 20 μL of proteinase K buffer (see below).
T19850	37218-37223	JJ	denotes	Whole
T19851	37224-37231	NNS	denotes	embryos
T19853	37232-37236	VBD	denotes	were
T19854	37237-37242	RB	denotes	first
T19852	37243-37251	VBN	denotes	digested
T19855	37252-37254	IN	denotes	in
T19856	37255-37257	CD	denotes	20
T19857	37258-37260	NN	denotes	μL
T19858	37261-37263	IN	denotes	of
T19859	37264-37274	NN	denotes	proteinase
T19860	37275-37276	NN	denotes	K
T19861	37277-37283	NN	denotes	buffer
T19862	37284-37285	-LRB-	denotes	(
T19863	37285-37288	VB	denotes	see
T19864	37289-37294	RB	denotes	below
T19865	37294-37295	-RRB-	denotes	)
T19866	37295-37296	.	denotes	.
T19867	37296-37473	sentence	denotes	The lysate was then divided in two, with half (10 μL) being used in the amplification of the wild-type allele and the remaining 10 μL in the amplification of the mutant allele.
T19868	37297-37300	DT	denotes	The
T19869	37301-37307	NN	denotes	lysate
T19871	37308-37311	VBD	denotes	was
T19872	37312-37316	RB	denotes	then
T19870	37317-37324	VBN	denotes	divided
T19873	37325-37327	IN	denotes	in
T19874	37328-37331	CD	denotes	two
T19875	37331-37333	,	denotes	,
T19876	37333-37337	IN	denotes	with
T19877	37338-37342	NN	denotes	half
T19879	37343-37344	-LRB-	denotes	(
T19881	37344-37346	CD	denotes	10
T19880	37347-37349	NN	denotes	μL
T19882	37349-37350	-RRB-	denotes	)
T19883	37351-37356	VBG	denotes	being
T19878	37357-37361	VBN	denotes	used
T19884	37362-37364	IN	denotes	in
T19885	37365-37368	DT	denotes	the
T19886	37369-37382	NN	denotes	amplification
T19887	37383-37385	IN	denotes	of
T19888	37386-37389	DT	denotes	the
T19890	37390-37394	JJ	denotes	wild
T19892	37394-37395	HYPH	denotes	-
T19891	37395-37399	NN	denotes	type
T19889	37400-37406	NN	denotes	allele
T19893	37407-37410	CC	denotes	and
T19894	37411-37414	DT	denotes	the
T19896	37415-37424	VBG	denotes	remaining
T19897	37425-37427	CD	denotes	10
T19895	37428-37430	NN	denotes	μL
T19898	37431-37433	IN	denotes	in
T19899	37434-37437	DT	denotes	the
T19900	37438-37451	NN	denotes	amplification
T19901	37452-37454	IN	denotes	of
T19902	37455-37458	DT	denotes	the
T19904	37459-37465	NN	denotes	mutant
T19903	37466-37472	NN	denotes	allele
T19905	37472-37473	.	denotes	.
T19906	37473-37677	sentence	denotes	The first reaction in the nested PCR amplification of the wild-type allele was carried out in a final reaction volume of 50 μL, using an intron 6 sense primer and an antisense primer located in intron 7.
T19907	37474-37477	DT	denotes	The
T19909	37478-37483	JJ	denotes	first
T19908	37484-37492	NN	denotes	reaction
T19911	37493-37495	IN	denotes	in
T19912	37496-37499	DT	denotes	the
T19914	37500-37506	VBN	denotes	nested
T19915	37507-37510	NN	denotes	PCR
T19913	37511-37524	NN	denotes	amplification
T19916	37525-37527	IN	denotes	of
T19917	37528-37531	DT	denotes	the
T19919	37532-37536	JJ	denotes	wild
T19921	37536-37537	HYPH	denotes	-
T19920	37537-37541	NN	denotes	type
T19918	37542-37548	NN	denotes	allele
T19922	37549-37552	VBD	denotes	was
T19910	37553-37560	VBN	denotes	carried
T19923	37561-37564	RP	denotes	out
T19924	37565-37567	IN	denotes	in
T19925	37568-37569	DT	denotes	a
T19927	37570-37575	JJ	denotes	final
T19928	37576-37584	NN	denotes	reaction
T19926	37585-37591	NN	denotes	volume
T19929	37592-37594	IN	denotes	of
T19930	37595-37597	CD	denotes	50
T19931	37598-37600	NN	denotes	μL
T19932	37600-37602	,	denotes	,
T19933	37602-37607	VBG	denotes	using
T19934	37608-37610	DT	denotes	an
T19936	37611-37617	NN	denotes	intron
T19937	37618-37619	CD	denotes	6
T19938	37620-37625	NN	denotes	sense
T19935	37626-37632	NN	denotes	primer
T19939	37633-37636	CC	denotes	and
T19940	37637-37639	DT	denotes	an
T19942	37640-37649	NN	denotes	antisense
T19941	37650-37656	NN	denotes	primer
T19943	37657-37664	VBN	denotes	located
T19944	37665-37667	IN	denotes	in
T19945	37668-37674	NN	denotes	intron
T19946	37675-37676	CD	denotes	7
T19947	37676-37677	.	denotes	.
T19948	37677-37831	sentence	denotes	Two μL of the first reaction were used as template in the second PCR amplification using another intron 6 sense primer and an antisense primer in exon 7.
T19949	37678-37681	CD	denotes	Two
T19950	37682-37684	NN	denotes	μL
T19952	37685-37687	IN	denotes	of
T19953	37688-37691	DT	denotes	the
T19955	37692-37697	JJ	denotes	first
T19954	37698-37706	NN	denotes	reaction
T19956	37707-37711	VBD	denotes	were
T19951	37712-37716	VBN	denotes	used
T19957	37717-37719	IN	denotes	as
T19958	37720-37728	NN	denotes	template
T19959	37729-37731	IN	denotes	in
T19960	37732-37735	DT	denotes	the
T19962	37736-37742	JJ	denotes	second
T19963	37743-37746	NN	denotes	PCR
T19961	37747-37760	NN	denotes	amplification
T19964	37761-37766	VBG	denotes	using
T19965	37767-37774	DT	denotes	another
T19967	37775-37781	NN	denotes	intron
T19968	37782-37783	CD	denotes	6
T19969	37784-37789	NN	denotes	sense
T19966	37790-37796	NN	denotes	primer
T19970	37797-37800	CC	denotes	and
T19971	37801-37803	DT	denotes	an
T19973	37804-37813	NN	denotes	antisense
T19972	37814-37820	NN	denotes	primer
T19974	37821-37823	IN	denotes	in
T19975	37824-37828	NN	denotes	exon
T19976	37829-37830	CD	denotes	7
T19977	37830-37831	.	denotes	.
T19978	37831-37904	sentence	denotes	The nested amplification of the mutant allele was carried out similarly.
T19979	37832-37835	DT	denotes	The
T19981	37836-37842	VBN	denotes	nested
T19980	37843-37856	NN	denotes	amplification
T19983	37857-37859	IN	denotes	of
T19984	37860-37863	DT	denotes	the
T19986	37864-37870	NN	denotes	mutant
T19985	37871-37877	NN	denotes	allele
T19987	37878-37881	VBD	denotes	was
T19982	37882-37889	VBN	denotes	carried
T19988	37890-37893	RP	denotes	out
T19989	37894-37903	RB	denotes	similarly
T19990	37903-37904	.	denotes	.
T19991	37904-38014	sentence	denotes	The first primer pair consisted of the intron 6 sense primer and an antisense primer in the PGK-Neo cassette.
T19992	37905-37908	DT	denotes	The
T19994	37909-37914	JJ	denotes	first
T19995	37915-37921	NN	denotes	primer
T19993	37922-37926	NN	denotes	pair
T19996	37927-37936	VBD	denotes	consisted
T19997	37937-37939	IN	denotes	of
T19998	37940-37943	DT	denotes	the
T20000	37944-37950	NN	denotes	intron
T20001	37951-37952	CD	denotes	6
T20002	37953-37958	NN	denotes	sense
T19999	37959-37965	NN	denotes	primer
T20003	37966-37969	CC	denotes	and
T20004	37970-37972	DT	denotes	an
T20006	37973-37982	NN	denotes	antisense
T20005	37983-37989	NN	denotes	primer
T20007	37990-37992	IN	denotes	in
T20008	37993-37996	DT	denotes	the
T20010	37997-38000	NN	denotes	PGK
T20012	38000-38001	HYPH	denotes	-
T20011	38001-38004	NN	denotes	Neo
T20009	38005-38013	NN	denotes	cassette
T20013	38013-38014	.	denotes	.
T20014	38014-38153	sentence	denotes	The nested primer pair was comprised of the second intron 6 sense primer and an antisense primer also located within the PGK-Neo sequence.
T20015	38015-38018	DT	denotes	The
T20017	38019-38025	VBN	denotes	nested
T20018	38026-38032	NN	denotes	primer
T20016	38033-38037	NN	denotes	pair
T20020	38038-38041	VBD	denotes	was
T20019	38042-38051	VBN	denotes	comprised
T20021	38052-38054	IN	denotes	of
T20022	38055-38058	DT	denotes	the
T20024	38059-38065	JJ	denotes	second
T20025	38066-38072	NN	denotes	intron
T20026	38073-38074	CD	denotes	6
T20027	38075-38080	NN	denotes	sense
T20023	38081-38087	NN	denotes	primer
T20028	38088-38091	CC	denotes	and
T20029	38092-38094	DT	denotes	an
T20031	38095-38104	NN	denotes	antisense
T20030	38105-38111	NN	denotes	primer
T20032	38112-38116	RB	denotes	also
T20033	38117-38124	VBN	denotes	located
T20034	38125-38131	IN	denotes	within
T20035	38132-38135	DT	denotes	the
T20037	38136-38139	NN	denotes	PGK
T20039	38139-38140	HYPH	denotes	-
T20038	38140-38143	NN	denotes	Neo
T20036	38144-38152	NN	denotes	sequence
T20040	38152-38153	.	denotes	.
T20041	38153-38283	sentence	denotes	It should be noted that amplification of both sequences involved the same sense primers in both steps of the nested PCR strategy.
T20042	38154-38156	PRP	denotes	It
T20044	38157-38163	MD	denotes	should
T20045	38164-38166	VB	denotes	be
T20043	38167-38172	VBN	denotes	noted
T20046	38173-38177	IN	denotes	that
T20048	38178-38191	NN	denotes	amplification
T20049	38192-38194	IN	denotes	of
T20050	38195-38199	DT	denotes	both
T20051	38200-38209	NNS	denotes	sequences
T20047	38210-38218	VBD	denotes	involved
T20052	38219-38222	DT	denotes	the
T20054	38223-38227	JJ	denotes	same
T20055	38228-38233	NN	denotes	sense
T20053	38234-38241	NNS	denotes	primers
T20056	38242-38244	IN	denotes	in
T20057	38245-38249	DT	denotes	both
T20058	38250-38255	NNS	denotes	steps
T20059	38256-38258	IN	denotes	of
T20060	38259-38262	DT	denotes	the
T20062	38263-38269	VBN	denotes	nested
T20063	38270-38273	NN	denotes	PCR
T20061	38274-38282	NN	denotes	strategy
T20064	38282-38283	.	denotes	.
T20065	38283-38499	sentence	denotes	In addition, the two sets of reactions included a common internal control designed to amplify a genomic region within the short arm of the targeting vector that is preserved in both the wild-type and mutant alleles.
T20066	38284-38286	IN	denotes	In
T20068	38287-38295	NN	denotes	addition
T20069	38295-38297	,	denotes	,
T20070	38297-38300	DT	denotes	the
T20072	38301-38304	CD	denotes	two
T20071	38305-38309	NNS	denotes	sets
T20073	38310-38312	IN	denotes	of
T20074	38313-38322	NNS	denotes	reactions
T20067	38323-38331	VBD	denotes	included
T20075	38332-38333	DT	denotes	a
T20077	38334-38340	JJ	denotes	common
T20078	38341-38349	JJ	denotes	internal
T20076	38350-38357	NN	denotes	control
T20079	38358-38366	VBN	denotes	designed
T20080	38367-38369	TO	denotes	to
T20081	38370-38377	VB	denotes	amplify
T20082	38378-38379	DT	denotes	a
T20084	38380-38387	JJ	denotes	genomic
T20083	38388-38394	NN	denotes	region
T20085	38395-38401	IN	denotes	within
T20086	38402-38405	DT	denotes	the
T20088	38406-38411	JJ	denotes	short
T20087	38412-38415	NN	denotes	arm
T20089	38416-38418	IN	denotes	of
T20090	38419-38422	DT	denotes	the
T20092	38423-38432	NN	denotes	targeting
T20091	38433-38439	NN	denotes	vector
T20093	38440-38444	WDT	denotes	that
T20095	38445-38447	VBZ	denotes	is
T20094	38448-38457	VBN	denotes	preserved
T20096	38458-38460	IN	denotes	in
T20097	38461-38465	CC	denotes	both
T20099	38466-38469	DT	denotes	the
T20100	38470-38474	JJ	denotes	wild
T20102	38474-38475	HYPH	denotes	-
T20101	38475-38479	NN	denotes	type
T20103	38480-38483	CC	denotes	and
T20104	38484-38490	NN	denotes	mutant
T20098	38491-38498	NNS	denotes	alleles
T20105	38498-38499	.	denotes	.
T20106	38499-38614	sentence	denotes	The first primer pair of the control PCR was made up of an intron 4 sense primer and an intron 5 antisense primer.
T20107	38500-38503	DT	denotes	The
T20109	38504-38509	JJ	denotes	first
T20110	38510-38516	NN	denotes	primer
T20108	38517-38521	NN	denotes	pair
T20112	38522-38524	IN	denotes	of
T20113	38525-38528	DT	denotes	the
T20115	38529-38536	NN	denotes	control
T20114	38537-38540	NN	denotes	PCR
T20116	38541-38544	VBD	denotes	was
T20111	38545-38549	VBN	denotes	made
T20117	38550-38552	RP	denotes	up
T20118	38553-38555	IN	denotes	of
T20119	38556-38558	DT	denotes	an
T20121	38559-38565	NN	denotes	intron
T20122	38566-38567	CD	denotes	4
T20123	38568-38573	NN	denotes	sense
T20120	38574-38580	NN	denotes	primer
T20124	38581-38584	CC	denotes	and
T20125	38585-38587	DT	denotes	an
T20127	38588-38594	NN	denotes	intron
T20128	38595-38596	CD	denotes	5
T20129	38597-38606	NN	denotes	antisense
T20126	38607-38613	NN	denotes	primer
T20130	38613-38614	.	denotes	.
T20131	38614-38710	sentence	denotes	The second primer pair comprised an exon 5 sense primer and a nested intron 5 antisense primer.
T20132	38615-38618	DT	denotes	The
T20134	38619-38625	JJ	denotes	second
T20135	38626-38632	NN	denotes	primer
T20133	38633-38637	NN	denotes	pair
T20136	38638-38647	VBD	denotes	comprised
T20137	38648-38650	DT	denotes	an
T20139	38651-38655	NN	denotes	exon
T20140	38656-38657	CD	denotes	5
T20141	38658-38663	NN	denotes	sense
T20138	38664-38670	NN	denotes	primer
T20142	38671-38674	CC	denotes	and
T20143	38675-38676	DT	denotes	a
T20145	38677-38683	VBN	denotes	nested
T20146	38684-38690	NN	denotes	intron
T20147	38691-38692	CD	denotes	5
T20148	38693-38702	NN	denotes	antisense
T20144	38703-38709	NN	denotes	primer
T20149	38709-38710	.	denotes	.
T20150	38710-38823	sentence	denotes	The final products were 213 bp for the control PCR, 429 bp for the wild-type PCR, and 389 bp for the mutant PCR.
T20151	38711-38714	DT	denotes	The
T20153	38715-38720	JJ	denotes	final
T20152	38721-38729	NNS	denotes	products
T20154	38730-38734	VBD	denotes	were
T20155	38735-38738	CD	denotes	213
T20156	38739-38741	NN	denotes	bp
T20157	38742-38745	IN	denotes	for
T20158	38746-38749	DT	denotes	the
T20160	38750-38757	NN	denotes	control
T20159	38758-38761	NN	denotes	PCR
T20161	38761-38763	,	denotes	,
T20162	38763-38766	CD	denotes	429
T20163	38767-38769	NN	denotes	bp
T20164	38770-38773	IN	denotes	for
T20165	38774-38777	DT	denotes	the
T20167	38778-38782	JJ	denotes	wild
T20169	38782-38783	HYPH	denotes	-
T20168	38783-38787	NN	denotes	type
T20166	38788-38791	NN	denotes	PCR
T20170	38791-38793	,	denotes	,
T20171	38793-38796	CC	denotes	and
T20172	38797-38800	CD	denotes	389
T20173	38801-38803	NN	denotes	bp
T20174	38804-38807	IN	denotes	for
T20175	38808-38811	DT	denotes	the
T20177	38812-38818	NN	denotes	mutant
T20176	38819-38822	NN	denotes	PCR
T20178	38822-38823	.	denotes	.
T20179	38823-39094	sentence	denotes	All reactions were carried out using identical PCR conditions entailing an initial five minute denaturation at 95°C, 35 cycles of one minute denaturation at 95°C, one minute annealing at 56°C, and one minute extension at 72°C, with a final extension step of ten minutes.
T20180	38824-38827	DT	denotes	All
T20181	38828-38837	NNS	denotes	reactions
T20183	38838-38842	VBD	denotes	were
T20182	38843-38850	VBN	denotes	carried
T20184	38851-38854	RP	denotes	out
T20185	38855-38860	VBG	denotes	using
T20186	38861-38870	JJ	denotes	identical
T20188	38871-38874	NN	denotes	PCR
T20187	38875-38885	NNS	denotes	conditions
T20189	38886-38895	VBG	denotes	entailing
T20190	38896-38898	DT	denotes	an
T20192	38899-38906	JJ	denotes	initial
T20193	38907-38911	CD	denotes	five
T20194	38912-38918	NN	denotes	minute
T20191	38919-38931	NN	denotes	denaturation
T20195	38932-38934	IN	denotes	at
T20196	38935-38937	CD	denotes	95
T20197	38937-38939	NN	denotes	°C
T20198	38939-38941	,	denotes	,
T20199	38941-38943	CD	denotes	35
T20200	38944-38950	NNS	denotes	cycles
T20201	38951-38953	IN	denotes	of
T20202	38954-38957	CD	denotes	one
T20204	38958-38964	NN	denotes	minute
T20203	38965-38977	NN	denotes	denaturation
T20205	38978-38980	IN	denotes	at
T20206	38981-38983	CD	denotes	95
T20207	38983-38985	NN	denotes	°C
T20208	38985-38987	,	denotes	,
T20209	38987-38990	CD	denotes	one
T20210	38991-38997	NN	denotes	minute
T20211	38998-39007	VBG	denotes	annealing
T20212	39008-39010	IN	denotes	at
T20213	39011-39013	CD	denotes	56
T20214	39013-39015	NN	denotes	°C
T20215	39015-39017	,	denotes	,
T20216	39017-39020	CC	denotes	and
T20217	39021-39024	CD	denotes	one
T20218	39025-39031	NN	denotes	minute
T20219	39032-39041	NN	denotes	extension
T20220	39042-39044	IN	denotes	at
T20221	39045-39047	CD	denotes	72
T20222	39047-39049	NN	denotes	°C
T20223	39049-39051	,	denotes	,
T20224	39051-39055	IN	denotes	with
T20225	39056-39057	DT	denotes	a
T20227	39058-39063	JJ	denotes	final
T20228	39064-39073	NN	denotes	extension
T20226	39074-39078	NN	denotes	step
T20229	39079-39081	IN	denotes	of
T20230	39082-39085	CD	denotes	ten
T20231	39086-39093	NNS	denotes	minutes
T20232	39093-39094	.	denotes	.
T20492	39096-39105	NN	denotes	Isolation
T20493	39106-39108	IN	denotes	of
T20494	39109-39125	JJ	denotes	pre-implantation
T20495	39126-39133	NNS	denotes	embryos
T20496	39133-39248	sentence	denotes	Time of fertilization was determined by observation of copulation plugs, and noon of that day was defined as E0.5.
T20497	39134-39138	NN	denotes	Time
T20499	39139-39141	IN	denotes	of
T20500	39142-39155	NN	denotes	fertilization
T20501	39156-39159	VBD	denotes	was
T20498	39160-39170	VBN	denotes	determined
T20502	39171-39173	IN	denotes	by
T20503	39174-39185	NN	denotes	observation
T20504	39186-39188	IN	denotes	of
T20505	39189-39199	NN	denotes	copulation
T20506	39200-39205	NNS	denotes	plugs
T20507	39205-39207	,	denotes	,
T20508	39207-39210	CC	denotes	and
T20509	39211-39215	NN	denotes	noon
T20511	39216-39218	IN	denotes	of
T20512	39219-39223	DT	denotes	that
T20513	39224-39227	NN	denotes	day
T20514	39228-39231	VBD	denotes	was
T20510	39232-39239	VBN	denotes	defined
T20515	39240-39242	IN	denotes	as
T20516	39243-39247	NN	denotes	E0.5
T20517	39247-39248	.	denotes	.
T20518	39248-39398	sentence	denotes	Pre-implantation embryos were obtained by dissecting the uteri out of pregnant females at E2.5 or E3.5 and flushing the oviducts with ES cell medium.
T20519	39249-39265	JJ	denotes	Pre-implantation
T20520	39266-39273	NNS	denotes	embryos
T20522	39274-39278	VBD	denotes	were
T20521	39279-39287	VBN	denotes	obtained
T20523	39288-39290	IN	denotes	by
T20524	39291-39301	VBG	denotes	dissecting
T20525	39302-39305	DT	denotes	the
T20526	39306-39311	NNS	denotes	uteri
T20527	39312-39315	IN	denotes	out
T20528	39316-39318	IN	denotes	of
T20529	39319-39327	JJ	denotes	pregnant
T20530	39328-39335	NNS	denotes	females
T20531	39336-39338	IN	denotes	at
T20532	39339-39343	NN	denotes	E2.5
T20533	39344-39346	CC	denotes	or
T20534	39347-39351	NN	denotes	E3.5
T20535	39352-39355	CC	denotes	and
T20536	39356-39364	VBG	denotes	flushing
T20537	39365-39368	DT	denotes	the
T20538	39369-39377	NNS	denotes	oviducts
T20539	39378-39382	IN	denotes	with
T20540	39383-39385	NN	denotes	ES
T20542	39386-39390	NN	denotes	cell
T20541	39391-39397	NN	denotes	medium
T20543	39397-39398	.	denotes	.
T20544	39398-39615	sentence	denotes	Isolated embryos were then digested for five hours at 55°C in 20 μL of lysis buffer (50 mM Tris-HCl, 0.5% Triton X-100, 200 μL/mL proteinase K, pH 8.0), followed by ten minutes at 95°C to inactivate the proteinase K.
T20545	39399-39407	VBN	denotes	Isolated
T20546	39408-39415	NNS	denotes	embryos
T20548	39416-39420	VBD	denotes	were
T20549	39421-39425	RB	denotes	then
T20547	39426-39434	VBN	denotes	digested
T20550	39435-39438	IN	denotes	for
T20551	39439-39443	CD	denotes	five
T20552	39444-39449	NNS	denotes	hours
T20553	39450-39452	IN	denotes	at
T20554	39453-39455	CD	denotes	55
T20555	39455-39457	NN	denotes	°C
T20556	39458-39460	IN	denotes	in
T20557	39461-39463	CD	denotes	20
T20558	39464-39466	NN	denotes	μL
T20559	39467-39469	IN	denotes	of
T20560	39470-39475	NN	denotes	lysis
T20561	39476-39482	NN	denotes	buffer
T20562	39483-39484	-LRB-	denotes	(
T20564	39484-39486	CD	denotes	50
T20565	39487-39489	NN	denotes	mM
T20567	39490-39494	NN	denotes	Tris
T20568	39494-39495	HYPH	denotes	-
T20566	39495-39498	NN	denotes	HCl
T20569	39498-39500	,	denotes	,
T20570	39500-39503	CD	denotes	0.5
T20571	39503-39504	NN	denotes	%
T20573	39505-39511	NNP	denotes	Triton
T20572	39512-39513	NN	denotes	X
T20574	39513-39514	HYPH	denotes	-
T20575	39514-39517	CD	denotes	100
T20576	39517-39519	,	denotes	,
T20577	39519-39522	CD	denotes	200
T20578	39523-39525	NN	denotes	μL
T20580	39525-39526	SYM	denotes	/
T20581	39526-39528	NN	denotes	mL
T20582	39529-39539	NN	denotes	proteinase
T20579	39540-39541	NN	denotes	K
T20583	39541-39543	,	denotes	,
T20563	39543-39545	NN	denotes	pH
T20584	39546-39549	CD	denotes	8.0
T20585	39549-39550	-RRB-	denotes	)
T20586	39550-39552	,	denotes	,
T20587	39552-39560	VBN	denotes	followed
T20588	39561-39563	IN	denotes	by
T20589	39564-39567	CD	denotes	ten
T20590	39568-39575	NNS	denotes	minutes
T20591	39576-39578	IN	denotes	at
T20592	39579-39581	CD	denotes	95
T20593	39581-39583	NN	denotes	°C
T20594	39584-39586	TO	denotes	to
T20595	39587-39597	VB	denotes	inactivate
T20596	39598-39601	DT	denotes	the
T20598	39602-39612	NN	denotes	proteinase
T20597	39613-39614	NN	denotes	K
T20599	39614-39615	.	denotes	.
T20600	39615-39658	sentence	denotes	Lysates were then used for PCR genotyping.
T20601	39616-39623	NNS	denotes	Lysates
T20603	39624-39628	VBD	denotes	were
T20604	39629-39633	RB	denotes	then
T20602	39634-39638	VBN	denotes	used
T20605	39639-39642	IN	denotes	for
T20606	39643-39646	NN	denotes	PCR
T20607	39647-39657	NN	denotes	genotyping
T20608	39657-39658	.	denotes	.
R10	T674	T673	amod	embryonic,development
R100	T774	T772	conj	both,one
R1000	T3995	T3994	pobj	release,in
R1001	T3996	T3995	prep	of,release
R1002	T3997	T3998	amod	focal,adhesions
R1003	T4061	T4059	punct	", ",mutation
R1004	T3998	T3996	pobj	adhesions,of
R1005	T3999	T4000	punct	[,9
R1006	T4000	T3985	parataxis	9,displayed
R1007	T4001	T4000	punct	],9
R1008	T4002	T3946	punct	.,cultured
R1009	T4062	T4063	dep	which,retained
R101	T775	T772	prep	of,one
R1010	T4004	T4005	det	An,knockout
R1011	T4005	T4009	nsubj	knockout,resulted
R1012	T4063	T4059	relcl	retained,mutation
R1013	T4006	T4007	advmod	independently,derived
R1014	T4007	T4005	amod	derived,knockout
R1015	T4008	T4005	compound	Capn4,knockout
R1016	T4064	T4065	det	some,level
R1017	T4010	T4005	punct	", ",knockout
R1018	T4011	T4005	prep	involving,knockout
R1019	T4065	T4063	dobj	level,retained
R102	T776	T777	det	these,enzymes
R1020	T4012	T4013	det	a,deletion
R1021	T4013	T4011	pobj	deletion,involving
R1022	T4014	T4015	advmod	more,extensive
R1023	T4066	T4065	amod	small,level
R1024	T4015	T4013	amod	extensive,deletion
R1025	T4016	T4013	prep	of,deletion
R1026	T4067	T4065	prep	of,level
R1027	T4017	T4018	det	the,gene
R1028	T4018	T4016	pobj	gene,of
R1029	T4019	T4009	punct	", ",resulted
R103	T777	T775	pobj	enzymes,of
R1030	T4020	T4009	prep	in,resulted
R1031	T4068	T4069	compound	calpain,activity
R1032	T4021	T4022	det	an,lethality
R1033	T4022	T4020	pobj	lethality,in
R1034	T4069	T4067	pobj	activity,of
R1035	T4023	T4022	amod	earlier,lethality
R1036	T4024	T4022	amod	embryonic,lethality
R1037	T4025	T4009	punct	", ",resulted
R1038	T4070	T4044	punct	", ",targeted
R1039	T4026	T4027	advmod	apparently,at
R104	T778	T770	prep	during,roles
R1040	T4027	T4009	prep	at,resulted
R1041	T4028	T4029	det	a,stage
R1042	T4029	T4027	pobj	stage,at
R1043	T4071	T4044	advcl	allowing,targeted
R1044	T4030	T4029	amod	pre-implantation,stage
R1045	T4031	T4032	punct	[,32
R1046	T4032	T4009	parataxis	32,resulted
R1047	T4072	T4071	prep	for,allowing
R1048	T4033	T4032	punct	],32
R1049	T4034	T4009	punct	.,resulted
R105	T779	T780	amod	mammalian,embryogenesis
R1050	T4073	T4074	poss	their,survival
R1051	T4036	T4037	det	The,times
R1052	T4037	T4039	nsubj	times,suggested
R1053	T4038	T4037	amod	different,times
R1054	T4040	T4037	prep	of,times
R1055	T4074	T4072	pobj	survival,for
R1056	T4041	T4042	amod	embryonic,lethality
R1057	T4042	T4040	pobj	lethality,of
R1058	T4043	T4044	mark	that,targeted
R1059	T4075	T4074	prep	to,survival
R106	T780	T778	pobj	embryogenesis,during
R1060	T4044	T4039	ccomp	targeted,suggested
R1061	T4045	T4046	det	the,mice
R1062	T4046	T4044	nsubjpass	mice,targeted
R1063	T4076	T4075	pobj	mid-gestation,to
R1064	T4047	T4046	amod	first,mice
R1065	T4048	T4046	amod	reported,mice
R1066	T4077	T4044	punct	", ",targeted
R1067	T4078	T4079	mark	while,represented
R1068	T4079	T4044	advcl	represented,targeted
R1069	T4080	T4081	det	the,mice
R107	T781	T745	punct	.,abolished
R1070	T4081	T4079	nsubj	mice,represented
R1071	T4082	T4081	amod	second,mice
R1072	T4083	T4081	amod	reported,mice
R1073	T4084	T4081	nmod	Capn4,mice
R1074	T4085	T4084	punct	-,Capn4
R1075	T4086	T4084	punct	/,Capn4
R1076	T4087	T4084	punct	-,Capn4
R1077	T4088	T4089	punct	[,32
R1078	T4049	T4046	nmod	Capn4,mice
R1079	T4089	T4081	parataxis	32,mice
R108	T783	T784	nsubj	Disruption,produced
R1080	T4090	T4089	punct	],32
R1081	T4050	T4049	punct	-,Capn4
R1082	T4091	T4092	det	a,mutation
R1083	T4051	T4049	punct	/,Capn4
R1084	T4052	T4049	punct	-,Capn4
R1085	T4092	T4079	dobj	mutation,represented
R1086	T4053	T4054	punct	[,31
R1087	T4054	T4046	parataxis	31,mice
R1088	T4093	T4092	amod	true,mutation
R1089	T4055	T4054	punct	],31
R109	T785	T783	prep	of,Disruption
R1090	T4094	T4092	amod	null,mutation
R1091	T4056	T4044	auxpass	were,targeted
R1092	T4057	T4044	prep	with,targeted
R1093	T4058	T4059	det	a,mutation
R1094	T4095	T4039	punct	.,suggested
R1095	T4059	T4057	pobj	mutation,with
R1096	T4060	T4059	amod	hypomorphic,mutation
R1097	T4097	T4098	nsubjpass	Disruption,reported
R1098	T4099	T4097	prep	of,Disruption
R1099	T4100	T4099	pobj	Capn1,of
R11	T675	T669	prep	in,required
R110	T786	T787	det	the,gene
R1100	T4101	T4097	punct	", ",Disruption
R1101	T4102	T4097	acl	encoding,Disruption
R1102	T4168	T4169	det	The,viability
R1103	T4169	T4170	nsubj	viability,distinguish
R1104	T4171	T4169	prep	of,viability
R1105	T4172	T4173	npadvmod	Capn1,deficient
R1106	T4173	T4175	amod	deficient,mice
R1107	T4174	T4173	punct	-,deficient
R1108	T4103	T4104	det	the,subunit
R1109	T4175	T4171	pobj	mice,of
R111	T787	T785	pobj	gene,of
R1110	T4176	T4170	aux	does,distinguish
R1111	T4177	T4170	neg	not,distinguish
R1112	T4104	T4102	dobj	subunit,encoding
R1113	T4178	T4170	advmod	however,distinguish
R1114	T4179	T4170	prep	between,distinguish
R1115	T4180	T4181	nummod	two,possibilities
R1116	T4105	T4106	nmod	μ,calpain
R1117	T4181	T4179	pobj	possibilities,between
R1118	T4182	T4181	punct	: ,possibilities
R1119	T4106	T4104	nmod	calpain,subunit
R112	T788	T787	compound	Capn1,gene
R1120	T4183	T4184	preconj	either,required
R1121	T4184	T4181	advcl	required,possibilities
R1122	T4185	T4184	mark	that,required
R1123	T4107	T4106	punct	-,calpain
R1124	T4186	T4187	compound	m,calpain
R1125	T4187	T4184	nsubjpass	calpain,required
R1126	T4188	T4187	punct	-,calpain
R1127	T4108	T4104	amod	catalytic,subunit
R1128	T4189	T4184	auxpass	is,required
R1129	T4190	T4184	advmod	specifically,required
R113	T789	T790	amod	viable,mice
R1130	T4109	T4098	punct	", ",reported
R1131	T4191	T4184	prep	during,required
R1132	T4192	T4191	pobj	embryogenesis,during
R1133	T4193	T4184	punct	", ",required
R1134	T4110	T4098	auxpass	was,reported
R1135	T4194	T4184	cc	or,required
R1136	T4195	T4196	mark	that,is
R1137	T4196	T4184	conj	is,required
R1138	T4111	T4098	advmod	subsequently,reported
R1139	T4197	T4198	preconj	either,form
R114	T790	T784	dobj	mice,produced
R1140	T4198	T4196	nsubj	form,is
R1141	T4199	T4198	prep	of,form
R1142	T4112	T4113	aux	to,result
R1143	T4200	T4199	pobj	calpain,of
R1144	T4201	T4198	advmod	alone,form
R1145	T4202	T4196	acomp	sufficient,is
R1146	T4113	T4098	xcomp	result,reported
R1147	T4203	T4196	cc	and,is
R1148	T4204	T4205	aux	can,compensate
R1149	T4114	T4113	prep	in,result
R115	T791	T790	punct	", ",mice
R1150	T4205	T4196	conj	compensate,is
R1151	T4206	T4205	prep	for,compensate
R1152	T4207	T4208	det	the,absence
R1153	T4115	T4116	amod	fertile,mice
R1154	T4208	T4206	pobj	absence,for
R1155	T4209	T4208	prep	of,absence
R1156	T4210	T4211	det	the,other
R1157	T4211	T4209	pobj	other,of
R1158	T4212	T4170	punct	.,distinguish
R1159	T4116	T4114	pobj	mice,in
R116	T792	T790	amod	fertile,mice
R1160	T4214	T4215	aux	To,resolve
R1161	T4215	T4216	advcl	resolve,knocked
R1162	T4117	T4116	punct	", ",mice
R1163	T4217	T4218	det	this,question
R1164	T4218	T4215	dobj	question,resolve
R1165	T4118	T4116	amod	viable,mice
R1166	T4219	T4216	punct	", ",knocked
R1167	T4220	T4216	nsubj	we,knocked
R1168	T4221	T4216	aux	have,knocked
R1169	T4119	T4116	prep	with,mice
R117	T793	T784	advcl	implying,produced
R1170	T4222	T4216	advmod	now,knocked
R1171	T4223	T4216	prt	out,knocked
R1172	T4224	T4225	det	the,gene
R1173	T4120	T4121	det	some,defects
R1174	T4225	T4216	dobj	gene,knocked
R1175	T4226	T4225	compound	Capn2,gene
R1176	T4227	T4225	acl	encoding,gene
R1177	T4121	T4119	pobj	defects,with
R1178	T4228	T4229	det	the,subunit
R1179	T4229	T4227	dobj	subunit,encoding
R118	T794	T795	mark	that,compensate
R1180	T4230	T4231	nmod	m,k
R1181	T4122	T4121	amod	mild,defects
R1182	T4231	T4229	compound	k,subunit
R1183	T4232	T4231	punct	-,k
R1184	T4233	T4231	nummod	80,k
R1185	T4123	T4121	prep	in,defects
R1186	T4234	T4227	prep	in,encoding
R1187	T4124	T4125	det	the,platelets
R1188	T4125	T4123	pobj	platelets,in
R1189	T4126	T4127	compound	μ,calpain
R119	T795	T793	advcl	compensate,implying
R1190	T4235	T4234	pobj	mice,in
R1191	T4236	T4216	punct	.,knocked
R1192	T4127	T4129	npadvmod	calpain,rich
R1193	T4238	T4239	nsubj	We,report
R1194	T4128	T4127	punct	-,calpain
R1195	T4240	T4239	advmod	here,report
R1196	T4241	T4242	mark	that,died
R1197	T4242	T4239	ccomp	died,report
R1198	T4129	T4125	amod	rich,platelets
R1199	T4243	T4244	npadvmod	Capn2,null
R12	T681	T682	compound	μ,calpain
R120	T796	T795	mark	either,compensate
R1200	T4244	T4245	amod	null,embryos
R1201	T4130	T4121	acl	relating,defects
R1202	T4245	T4242	nsubj	embryos,died
R1203	T4246	T4247	amod	prior,to
R1204	T4131	T4130	prep	to,relating
R1205	T4247	T4242	prep	to,died
R1206	T4248	T4249	det	the,stage
R1207	T4249	T4247	pobj	stage,to
R1208	T4250	T4249	compound	implantation,stage
R1209	T4251	T4242	punct	", ",died
R121	T797	T798	compound	m,calpain
R1210	T4252	T4242	advcl	indicating,died
R1211	T4132	T4133	poss	their,aggregation
R1212	T4253	T4254	mark	that,is
R1213	T4254	T4252	ccomp	is,indicating
R1214	T4255	T4256	compound	m,calpain
R1215	T4133	T4131	pobj	aggregation,to
R1216	T4256	T4254	nsubj	calpain,is
R1217	T4257	T4256	punct	-,calpain
R1218	T4258	T4254	acomp	indispensable,is
R1219	T4134	T4133	cc	and,aggregation
R122	T798	T795	nsubj	calpain,compensate
R1220	T4259	T4258	prep	for,indispensable
R1221	T4260	T4261	amod	early,embryogenesis
R1222	T4261	T4259	pobj	embryogenesis,for
R1223	T4135	T4136	compound	clot,retraction
R1224	T4262	T4239	punct	.,report
R1225	T4264	T4265	det	This,role
R1226	T4136	T4133	conj	retraction,aggregation
R1227	T4265	T4266	nsubjpass	role,fulfilled
R1228	T4267	T4266	aux	can,fulfilled
R1229	T4137	T4138	punct	[,33
R123	T799	T798	punct	-,calpain
R1230	T4268	T4266	neg	not,fulfilled
R1231	T4269	T4266	auxpass	be,fulfilled
R1232	T4270	T4266	agent	by,fulfilled
R1233	T4138	T4098	parataxis	33,reported
R1234	T4271	T4272	compound	μ,calpain
R1235	T4272	T4270	pobj	calpain,by
R1236	T4139	T4138	punct	],33
R1237	T4140	T4098	punct	.,reported
R1238	T4142	T4143	det	The,fact
R1239	T4273	T4272	punct	-,calpain
R124	T800	T795	aux	could,compensate
R1240	T4274	T4272	punct	", ",calpain
R1241	T4275	T4276	dep	which,expressed
R1242	T4143	T4144	nsubj	fact,indicates
R1243	T4276	T4272	relcl	expressed,calpain
R1244	T4277	T4276	auxpass	is,expressed
R1245	T4278	T4276	prep	in,expressed
R1246	T4145	T4146	mark	that,die
R1247	T4279	T4280	amod	embryonic,stem
R1248	T4280	T4281	nmod	stem,cells
R1249	T4281	T4278	pobj	cells,in
R125	T801	T795	prep	for,compensate
R1250	T4282	T4280	punct	(,stem
R1251	T4146	T4143	acl	die,fact
R1252	T4283	T4280	appos	ES,stem
R1253	T4284	T4281	punct	),cells
R1254	T4285	T4286	punct	[,31
R1255	T4147	T4148	npadvmod	Capn4,null
R1256	T4286	T4276	parataxis	31,expressed
R1257	T4287	T4286	punct	],31
R1258	T4288	T4276	cc	and,expressed
R1259	T4148	T4149	amod	null,mice
R126	T802	T803	det	the,loss
R1260	T4289	T4290	auxpass	is,assumed
R1261	T4290	T4276	conj	assumed,expressed
R1262	T4291	T4292	aux	to,be
R1263	T4292	T4290	xcomp	be,assumed
R1264	T4293	T4292	acomp	present,be
R1265	T4294	T4293	prep	at,present
R1266	T4149	T4146	nsubj	mice,die
R1267	T4295	T4296	det	this,stage
R1268	T4296	T4294	pobj	stage,at
R1269	T4297	T4296	prep	of,stage
R127	T803	T801	pobj	loss,for
R1270	T4150	T4146	prep	during,die
R1271	T4298	T4297	pobj	gestation,of
R1272	T4299	T4266	punct	.,fulfilled
R1273	T4151	T4150	pobj	embryogenesis,during
R1274	T4301	T4302	nsubj	This,demonstrates
R1275	T4152	T4153	mark	that,is
R1276	T4303	T4302	advmod	unequivocally,demonstrates
R1277	T4304	T4305	mark	that,have
R1278	T4305	T4302	ccomp	have,demonstrates
R1279	T4153	T4144	ccomp	is,indicates
R128	T804	T803	prep	of,loss
R1280	T4306	T4307	compound	m,calpain
R1281	T4307	T4305	nsubj	calpain,have
R1282	T4308	T4307	punct	-,calpain
R1283	T4154	T4155	advmod	at,one
R1284	T4309	T4307	cc	and,calpain
R1285	T4310	T4311	compound	μ,calpain
R1286	T4311	T4307	conj	calpain,calpain
R1287	T4155	T4153	nsubj	one,is
R1288	T4312	T4311	punct	-,calpain
R1289	T4313	T4314	amod	distinct,roles
R129	T805	T806	compound	μ,calpain
R1290	T4314	T4305	dobj	roles,have
R1291	T4156	T4155	advmod	least,one
R1292	T4315	T4314	amod	physiological,roles
R1293	T4316	T4305	prep	during,have
R1294	T4317	T4318	amod	early,embryogenesis
R1295	T4157	T4155	prep	of,one
R1296	T4318	T4316	pobj	embryogenesis,during
R1297	T4319	T4302	punct	.,demonstrates
R1298	T4158	T4159	det	the,calpains
R1299	T4159	T4157	pobj	calpains,of
R13	T682	T684	nsubj	calpain,are
R130	T806	T804	pobj	calpain,of
R1300	T4160	T4159	amod	ubiquitous,calpains
R1301	T4161	T4153	acomp	essential,is
R1302	T4162	T4161	prep	for,essential
R1303	T4163	T4162	pobj	development,for
R1304	T4164	T4163	prep	to,development
R1305	T4165	T4164	pobj	term,to
R1306	T4166	T4144	punct	.,indicates
R1307	T4833	T4832	cc	and,Isolation
R1308	T4834	T4832	conj	characterization,Isolation
R1309	T4835	T4832	prep	of,Isolation
R131	T807	T806	punct	-,calpain
R1310	T4836	T4837	npadvmod	Capn2,targeted
R1311	T4837	T4838	amod	targeted,clones
R1312	T4838	T4835	pobj	clones,of
R1313	T4839	T4840	compound	ES,cell
R1314	T4840	T4838	compound	cell,clones
R1315	T4842	T4843	nummod	Two,lines
R1316	T4843	T4851	nsubjpass	lines,isolated
R1317	T4844	T4843	amod	independent,lines
R1318	T4845	T4843	nmod	Capn2,lines
R1319	T4846	T4845	punct	+,Capn2
R132	T808	T795	punct	", ",compensate
R1320	T4847	T4845	punct	/,Capn2
R1321	T4848	T4845	punct	-,Capn2
R1322	T4849	T4843	compound	ES,lines
R1323	T4850	T4843	compound	cell,lines
R1324	T4852	T4843	punct	", ",lines
R1325	T4853	T4843	acl	designated,lines
R1326	T4854	T4853	oprd	ES27,designated
R1327	T4855	T4854	cc	and,ES27
R1328	T4856	T4854	conj	ES36,ES27
R1329	T4857	T4851	punct	", ",isolated
R133	T809	T795	cc	or,compensate
R1330	T4858	T4851	auxpass	were,isolated
R1331	T4859	T4851	prep	from,isolated
R1332	T4860	T4861	det	a,screen
R1333	T4861	T4859	pobj	screen,from
R1334	T4862	T4861	prep	of,screen
R1335	T4863	T4864	nummod	305,clones
R1336	T4864	T4862	pobj	clones,of
R1337	T4865	T4866	npadvmod	drug,resistant
R1338	T4866	T4864	amod	resistant,clones
R1339	T4867	T4866	punct	-,resistant
R134	T810	T811	mark	that,was
R1340	T4868	T4851	punct	.,isolated
R1341	T4870	T4871	amod	Correct,targeting
R1342	T4871	T4872	nsubjpass	targeting,established
R1343	T4873	T4871	prep	of,targeting
R1344	T4874	T4875	det	the,locus
R1345	T4875	T4873	pobj	locus,of
R1346	T4876	T4875	compound	Capn2,locus
R1347	T4877	T4872	auxpass	was,established
R1348	T4878	T4879	preconj	both,by
R1349	T4879	T4872	agent	by,established
R135	T811	T795	conj	was,compensate
R1350	T4880	T4881	compound	Southern,blot
R1351	T4881	T4882	compound	blot,hybridization
R1352	T4882	T4879	pobj	hybridization,by
R1353	T4883	T4882	cc	and,hybridization
R1354	T4884	T4885	compound	PCR,analysis
R1355	T4885	T4882	conj	analysis,hybridization
R1356	T4886	T4872	punct	.,established
R1357	T4888	T4889	det	A,probe
R1358	T4889	T4890	nsubj	probe,hybridized
R1359	T4891	T4889	acl	located,probe
R136	T812	T813	det	the,loss
R1360	T4892	T4891	advmod	outside,located
R1361	T4893	T4894	det	the,arm
R1362	T4894	T4892	npadvmod	arm,outside
R1363	T4895	T4894	amod	short,arm
R1364	T4896	T4897	punct	(,upstream
R1365	T4897	T4895	parataxis	upstream,short
R1366	T4898	T4897	punct	),upstream
R1367	T4899	T4894	prep	of,arm
R1368	T4900	T4899	pobj	homology,of
R1369	T4901	T4890	prep	to,hybridized
R137	T813	T811	nsubj	loss,was
R1370	T4902	T4903	det	a,fragment
R1371	T4903	T4901	pobj	fragment,to
R1372	T4904	T4905	nummod	3.5,kb
R1373	T4905	T4903	compound	kb,fragment
R1374	T4906	T4905	punct	-,kb
R1375	T4907	T4903	compound	BamHI,fragment
R1376	T4908	T4903	prep	of,fragment
R1377	T4909	T4910	det	the,allele
R1378	T4910	T4908	pobj	allele,of
R1379	T4911	T4912	amod	wild,type
R138	T814	T813	prep	of,loss
R1380	T4912	T4910	compound	type,allele
R1381	T4913	T4912	punct	-,type
R1382	T4914	T4903	cc	and,fragment
R1383	T4915	T4916	nummod	5.3,kb
R1384	T4916	T4918	compound	kb,fragment
R1385	T4917	T4916	punct	-,kb
R1386	T4918	T4903	conj	fragment,fragment
R1387	T4919	T4918	compound	BamHI,fragment
R1388	T4920	T4918	prep	of,fragment
R1389	T4921	T4922	det	the,allele
R139	T815	T816	compound	m,calpain
R1390	T4922	T4920	pobj	allele,of
R1391	T4923	T4922	compound	mutant,allele
R1392	T4924	T4925	mark	as,predicted
R1393	T4925	T4890	advcl	predicted,hybridized
R1394	T4926	T4925	prep	from,predicted
R1395	T4927	T4928	amod	genomic,maps
R1396	T4928	T4926	pobj	maps,from
R1397	T4929	T4930	punct	(,2A
R1398	T4930	T4925	parataxis	2A,predicted
R1399	T4931	T4930	compound	Figure,2A
R14	T683	T682	punct	-,calpain
R140	T816	T814	pobj	calpain,of
R1400	T4932	T4930	punct	),2A
R1401	T4933	T4890	punct	.,hybridized
R1402	T4935	T4936	det	The,probe
R1403	T4936	T4938	nsubj	probe,detected
R1404	T4937	T4936	amod	same,probe
R1405	T4939	T4938	advmod	also,detected
R1406	T4940	T4941	det	the,fragments
R1407	T4941	T4938	dobj	fragments,detected
R1408	T4942	T4941	amod	expected,fragments
R1409	T4943	T4944	nummod	7.2,kb
R141	T817	T816	punct	-,calpain
R1410	T4944	T4941	nmod	kb,fragments
R1411	T4945	T4944	punct	-,kb
R1412	T4946	T4947	amod	wild,type
R1413	T4947	T4944	appos	type,kb
R1414	T4948	T4947	punct	-,type
R1415	T4949	T4944	cc	and,kb
R1416	T4950	T4951	nummod	6.4,kb
R1417	T4951	T4953	compound	kb,mutant
R1418	T4952	T4951	punct	-,kb
R1419	T4953	T4944	conj	mutant,kb
R142	T818	T811	acomp	responsible,was
R1420	T4954	T4941	compound	BglII,fragments
R1421	T4955	T4941	punct	", ",fragments
R1422	T4956	T4957	nummod	4.9,kb
R1423	T4957	T4959	nmod	kb,fragments
R1424	T4958	T4957	punct	-,kb
R1425	T4959	T4941	conj	fragments,fragments
R1426	T4960	T4961	amod	wild,type
R1427	T4961	T4957	appos	type,kb
R1428	T4962	T4961	punct	-,type
R1429	T4963	T4957	cc	and,kb
R143	T819	T818	prep	for,responsible
R1430	T4964	T4965	nummod	5.7,kb
R1431	T4965	T4967	compound	kb,mutant
R1432	T4966	T4965	punct	-,kb
R1433	T4967	T4957	conj	mutant,kb
R1434	T4968	T4959	compound	NcoI,fragments
R1435	T4969	T4959	punct	", ",fragments
R1436	T4970	T4971	advmod	as,as
R1437	T4971	T4959	cc	as,fragments
R1438	T4972	T4971	advmod	well,as
R1439	T4973	T4974	nummod	7.2,kb
R144	T820	T819	pobj	death,for
R1440	T4974	T4976	nmod	kb,fragments
R1441	T4975	T4974	punct	-,kb
R1442	T4976	T4959	conj	fragments,fragments
R1443	T4977	T4978	amod	wild,type
R1444	T4978	T4974	appos	type,kb
R1445	T4979	T4978	punct	-,type
R1446	T4980	T4974	cc	and,kb
R1447	T4981	T4982	nummod	4.9,kb
R1448	T4982	T4984	compound	kb,mutant
R1449	T4983	T4982	punct	-,kb
R145	T821	T820	prep	of,death
R1450	T4984	T4974	conj	mutant,kb
R1451	T4985	T4986	compound	BglII,AgeI
R1452	T4986	T4976	compound	AgeI,fragments
R1453	T4987	T4986	punct	/,AgeI
R1454	T4988	T4989	punct	(,shown
R1455	T4989	T4938	parataxis	shown,detected
R1456	T4990	T4989	neg	not,shown
R1457	T4991	T4989	punct	),shown
R1458	T4992	T4938	punct	.,detected
R1459	T4994	T4995	det	A,probe
R146	T822	T823	nmod	Capn4,mice
R1460	T4995	T4996	nsubj	probe,recognized
R1461	T4997	T4995	acl	derived,probe
R1462	T4998	T4997	prep	from,derived
R1463	T4999	T5000	det	the,cassette
R1464	T5000	T4998	pobj	cassette,from
R1465	T5001	T5002	compound	PGK,Neo
R1466	T5002	T5000	compound	Neo,cassette
R1467	T5003	T5002	punct	-,Neo
R1468	T5004	T5005	advmod	only,fragment
R1469	T5005	T4996	dobj	fragment,recognized
R147	T823	T821	pobj	mice,of
R1470	T5006	T5005	det	the,fragment
R1471	T5007	T5008	nummod	5.3,kb
R1472	T5008	T5005	compound	kb,fragment
R1473	T5009	T5008	punct	-,kb
R1474	T5010	T5005	compound	BamHI,fragment
R1475	T5011	T4996	prep	in,recognized
R1476	T5012	T5013	nmod	Capn2,ES
R1477	T5013	T5017	compound	ES,cells
R1478	T5014	T5013	punct	+,ES
R1479	T5015	T5013	punct	/,ES
R148	T824	T822	punct	-,Capn4
R1480	T5016	T5013	punct	-,ES
R1481	T5017	T5011	pobj	cells,in
R1482	T5018	T4996	punct	", ",recognized
R1483	T5019	T4996	advcl	suggesting,recognized
R1484	T5020	T5021	mark	that,integrated
R1485	T5021	T5019	ccomp	integrated,suggesting
R1486	T5022	T5023	det	the,vector
R1487	T5023	T5021	nsubj	vector,integrated
R1488	T5024	T5023	compound	targeting,vector
R1489	T5025	T5021	aux	had,integrated
R149	T825	T822	punct	/,Capn4
R1490	T5026	T5027	advmod	solely,at
R1491	T5027	T5021	prep	at,integrated
R1492	T5028	T5029	det	the,locus
R1493	T5029	T5027	pobj	locus,at
R1494	T5030	T5029	compound	Capn2,locus
R1495	T5031	T5032	punct	(,shown
R1496	T5032	T5019	parataxis	shown,suggesting
R1497	T5033	T5032	neg	not,shown
R1498	T5034	T5032	punct	),shown
R1499	T5035	T4996	punct	.,recognized
R15	T685	T682	cc	and,calpain
R150	T826	T822	punct	-,Capn4
R1500	T5037	T5038	det	A,method
R1501	T5038	T5041	nsubjpass	method,established
R1502	T5039	T5038	compound	PCR,method
R1503	T5040	T5038	compound	screening,method
R1504	T5042	T5041	auxpass	was,established
R1505	T5043	T5041	advmod	also,established
R1506	T5044	T5045	dep	that,generated
R1507	T5045	T5041	ccomp	generated,established
R1508	T5046	T5047	det	a,product
R1509	T5047	T5045	dobj	product,generated
R151	T827	T784	punct	.,produced
R1510	T5048	T5049	amod	wild,type
R1511	T5049	T5047	compound	type,product
R1512	T5050	T5049	punct	-,type
R1513	T5051	T5047	prep	of,product
R1514	T5052	T5053	nummod	"2,749",bp
R1515	T5053	T5051	pobj	bp,of
R1516	T5054	T5047	cc	and,product
R1517	T5055	T5056	det	a,product
R1518	T5056	T5047	conj	product,product
R1519	T5057	T5058	nummod	"2,711",bp
R152	T831	T832	aux	To,distinguish
R1520	T5058	T5056	compound	bp,product
R1521	T5059	T5056	prep	from,product
R1522	T5060	T5061	det	the,allele
R1523	T5061	T5059	pobj	allele,from
R1524	T5062	T5061	compound	mutant,allele
R1525	T5063	T5041	punct	.,established
R1526	T5065	T5066	det	The,product
R1527	T5066	T5069	nsubj	product,was
R1528	T5067	T5068	nummod	"2,711",bp
R1529	T5068	T5066	compound	bp,product
R153	T832	T833	advcl	distinguish,deleted
R1530	T5070	T5069	advmod	only,was
R1531	T5071	T5069	acomp	evident,was
R1532	T5072	T5069	prep	in,was
R1533	T5073	T5074	det	the,lines
R1534	T5074	T5072	pobj	lines,in
R1535	T5075	T5074	nummod	two,lines
R1536	T5076	T5074	amod	targeted,lines
R1537	T5077	T5074	compound	cell,lines
R1538	T5078	T5079	punct	(,2B
R1539	T5079	T5069	parataxis	2B,was
R154	T834	T832	prep	between,distinguish
R1540	T5080	T5079	compound	Figure,2B
R1541	T5081	T5079	punct	),2B
R1542	T5082	T5069	punct	.,was
R1543	T6636	T6635	prep	of,Generation
R1544	T6637	T6638	amod	chimeric,mice
R1545	T6638	T6636	pobj	mice,of
R1546	T6639	T6635	cc	and,Generation
R1547	T6640	T6641	compound	germline,transmission
R1548	T6641	T6635	conj	transmission,Generation
R1549	T6642	T6641	prep	of,transmission
R155	T835	T836	det	the,alternatives
R1550	T6643	T6644	det	the,allele
R1551	T6644	T6642	pobj	allele,of
R1552	T6645	T6644	compound	mutant,allele
R1553	T6646	T6644	compound	Capn2,allele
R1554	T6648	T6649	nummod	Eight,mice
R1555	T6649	T6652	nsubjpass	mice,produced
R1556	T6650	T6649	amod	chimeric,mice
R1557	T6651	T6649	amod	male,mice
R1558	T6653	T6652	auxpass	were,produced
R1559	T6654	T6652	prep	in,produced
R156	T836	T834	pobj	alternatives,between
R1560	T6655	T6656	compound	morula,aggregation
R1561	T6656	T6657	compound	aggregation,experiments
R1562	T6657	T6654	pobj	experiments,in
R1563	T6658	T6652	advcl	using,produced
R1564	T6659	T6660	det	the,line
R1565	T6660	T6658	dobj	line,using
R1566	T6661	T6660	nmod	Capn2,line
R1567	T6662	T6661	punct	+,Capn2
R1568	T6663	T6661	punct	/,Capn2
R1569	T6664	T6661	punct	-,Capn2
R157	T837	T836	acl	described,alternatives
R1570	T6665	T6660	compound	ES27,line
R1571	T6666	T6660	compound	cell,line
R1572	T6667	T6652	punct	.,produced
R1573	T6669	T6670	nsubj	Two,transmitted
R1574	T6671	T6669	prep	of,Two
R1575	T6672	T6673	det	these,males
R1576	T6673	T6671	pobj	males,of
R1577	T6674	T6675	det	the,genotype
R1578	T6675	T6670	dobj	genotype,transmitted
R1579	T6676	T6675	nmod	Capn2,genotype
R158	T838	T837	advmod	above,described
R1580	T6677	T6676	punct	+,Capn2
R1581	T6678	T6676	punct	/,Capn2
R1582	T6679	T6676	punct	-,Capn2
R1583	T6680	T6675	compound	ES27,genotype
R1584	T6681	T6670	prep	through,transmitted
R1585	T6682	T6683	det	the,germline
R1586	T6683	T6681	pobj	germline,through
R1587	T6684	T6670	prep	into,transmitted
R1588	T6685	T6686	det	the,generation
R1589	T6686	T6684	pobj	generation,into
R159	T839	T833	punct	", ",deleted
R1590	T6687	T6686	compound	F1,generation
R1591	T6688	T6670	punct	.,transmitted
R1592	T6690	T6691	amod	Heterozygous,animals
R1593	T6691	T6696	nsubj	animals,appeared
R1594	T6692	T6691	nmod	Capn2,animals
R1595	T6693	T6692	punct	+,Capn2
R1596	T6694	T6692	punct	/,Capn2
R1597	T6695	T6692	punct	-,Capn2
R1598	T6697	T6696	oprd	normal,appeared
R1599	T6698	T6696	punct	", ",appeared
R16	T686	T687	compound	m,calpain
R160	T840	T833	nsubj	we,deleted
R1600	T6699	T6696	prep	with,appeared
R1601	T6700	T6701	det	no,defects
R1602	T6701	T6699	pobj	defects,with
R1603	T6702	T6701	amod	obvious,defects
R1604	T6703	T6701	prep	in,defects
R1605	T6704	T6705	amod	gross,anatomy
R1606	T6705	T6703	pobj	anatomy,in
R1607	T6706	T6705	punct	", ",anatomy
R1608	T6707	T6705	conj	reproduction,anatomy
R1609	T6708	T6707	punct	", ",reproduction
R161	T841	T842	det	an,region
R1610	T6709	T6707	cc	or,reproduction
R1611	T6710	T6711	compound	life,span
R1612	T6711	T6707	conj	span,reproduction
R1613	T6712	T6696	punct	.,appeared
R1614	T6714	T6715	prep	Out,detected
R1615	T6716	T6714	prep	of,Out
R1616	T6717	T6718	nummod	199,weanlings
R1617	T6718	T6716	pobj	weanlings,of
R1618	T6719	T6718	prep	from,weanlings
R1619	T6720	T6721	amod	heterozygous,intercrosses
R162	T842	T833	dobj	region,deleted
R1620	T6721	T6719	pobj	intercrosses,from
R1621	T6722	T6715	punct	", ",detected
R1622	T6723	T6724	det	no,progeny
R1623	T6724	T6715	nsubjpass	progeny,detected
R1624	T6725	T6724	nmod	Capn2,progeny
R1625	T6726	T6725	punct	-,Capn2
R1626	T6727	T6725	punct	/,Capn2
R1627	T6728	T6725	punct	-,Capn2
R1628	T6729	T6715	auxpass	were,detected
R1629	T6730	T6731	punct	(,Table
R163	T843	T842	amod	essential,region
R1630	T6731	T6715	parataxis	Table,detected
R1631	T6732	T6731	nummod	2,Table
R1632	T6733	T6731	punct	),Table
R1633	T6734	T6715	punct	.,detected
R1634	T6736	T6737	nsubj	We,observe
R1635	T6738	T6737	aux	did,observe
R1636	T6739	T6737	neg	not,observe
R1637	T6740	T6741	amod	high,rates
R1638	T6741	T6737	dobj	rates,observe
R1639	T6742	T6741	prep	of,rates
R164	T844	T842	compound	coding,region
R1640	T6743	T6744	amod	perinatal,death
R1641	T6744	T6742	pobj	death,of
R1642	T6745	T6737	punct	", ",observe
R1643	T6746	T6737	cc	and,observe
R1644	T6747	T6748	det	no,stillborns
R1645	T6748	T6753	nsubjpass	stillborns,observed
R1646	T6749	T6748	nmod	Capn2,stillborns
R1647	T6750	T6749	punct	-,Capn2
R1648	T6751	T6749	punct	/,Capn2
R1649	T6752	T6749	punct	-,Capn2
R165	T845	T842	prep	in,region
R1650	T6753	T6737	conj	observed,observe
R1651	T6754	T6753	auxpass	were,observed
R1652	T6755	T6753	punct	.,observed
R1653	T6757	T6758	nsubj	This,suggested
R1654	T6759	T6760	mark	that,perished
R1655	T6760	T6758	ccomp	perished,suggested
R1656	T6761	T6762	nmod	Capn2,animals
R1657	T6762	T6760	nsubj	animals,perished
R1658	T6763	T6761	punct	-,Capn2
R1659	T6764	T6761	punct	/,Capn2
R166	T846	T847	det	the,gene
R1660	T6765	T6761	punct	-,Capn2
R1661	T6766	T6760	prep	at,perished
R1662	T6767	T6768	det	some,stage
R1663	T6768	T6766	pobj	stage,at
R1664	T6769	T6768	prep	during,stage
R1665	T6770	T6771	amod	embryonic,development
R1666	T6771	T6769	pobj	development,during
R1667	T6772	T6758	punct	.,suggested
R1668	T6774	T6775	prep	In,harvested
R1669	T6776	T6777	det	an,attempt
R167	T847	T845	pobj	gene,in
R1670	T6777	T6774	pobj	attempt,In
R1671	T6778	T6779	aux	to,determine
R1672	T6779	T6777	acl	determine,attempt
R1673	T6780	T6781	mark	if,occurred
R1674	T6781	T6779	ccomp	occurred,determine
R1675	T6782	T6783	amod	embryonic,death
R1676	T6783	T6781	nsubj	death,occurred
R1677	T6784	T6781	prep	at,occurred
R1678	T6785	T6786	det	a,stage
R1679	T6786	T6784	pobj	stage,at
R168	T848	T847	compound	mouse,gene
R1680	T6787	T6786	amod	post-implantation,stage
R1681	T6788	T6775	punct	", ",harvested
R1682	T6789	T6775	nsubjpass	embryos,harvested
R1683	T6790	T6775	auxpass	were,harvested
R1684	T6791	T6775	prep	for,harvested
R1685	T6792	T6791	pobj	genotyping,for
R1686	T6793	T6775	prep	at,harvested
R1687	T6794	T6795	amod	different,times
R1688	T6795	T6793	pobj	times,at
R1689	T6796	T6795	prep	between,times
R169	T849	T847	compound	Capn2,gene
R1690	T6797	T6796	pobj	E10.5,between
R1691	T6798	T6797	cc	and,E10.5
R1692	T6799	T6797	conj	E18.5,E10.5
R1693	T6800	T6775	punct	.,harvested
R1694	T6802	T6803	det	No,embryos
R1695	T6803	T6808	nsubjpass	embryos,observed
R1696	T6804	T6803	nmod	Capn2,embryos
R1697	T6805	T6804	punct	-,Capn2
R1698	T6806	T6804	punct	/,Capn2
R1699	T6807	T6804	punct	-,Capn2
R17	T687	T682	conj	calpain,calpain
R170	T850	T833	prep	in,deleted
R1700	T6809	T6808	auxpass	were,observed
R1701	T6810	T6808	cc	and,observed
R1702	T6811	T6812	det	no,signs
R1703	T6812	T6813	nsubjpass	signs,detected
R1704	T6813	T6808	conj	detected,observed
R1705	T6814	T6812	prep	of,signs
R1706	T6815	T6816	compound	embryo,resorption
R1707	T6816	T6814	pobj	resorption,of
R1708	T6817	T6813	auxpass	were,detected
R1709	T6818	T6819	punct	(,Table
R171	T851	T852	amod	embryonic,cells
R1710	T6819	T6813	parataxis	Table,detected
R1711	T6820	T6819	nummod	2,Table
R1712	T6821	T6819	punct	),Table
R1713	T6822	T6813	punct	.,detected
R1714	T6824	T6825	nsubj	This,indicated
R1715	T6826	T6827	mark	that,dying
R1716	T6827	T6825	ccomp	dying,indicated
R1717	T6828	T6829	det	the,embryos
R1718	T6829	T6827	nsubj	embryos,dying
R1719	T6830	T6829	nmod	Capn2,embryos
R172	T852	T850	pobj	cells,in
R1720	T6831	T6830	punct	-,Capn2
R1721	T6832	T6830	punct	/,Capn2
R1722	T6833	T6830	punct	-,Capn2
R1723	T6834	T6827	aux	might,dying
R1724	T6835	T6827	aux	be,dying
R1725	T6836	T6837	amod	prior,to
R1726	T6837	T6827	prep	to,dying
R1727	T6838	T6837	pobj	implantation,to
R1728	T6839	T6825	punct	.,indicated
R1729	T6841	T6842	nsubjpass	Embryos,flushed
R173	T853	T852	nmod	stems,cells
R1730	T6843	T6842	auxpass	were,flushed
R1731	T6844	T6842	advmod	then,flushed
R1732	T6845	T6842	prep	from,flushed
R1733	T6846	T6847	det	the,oviducts
R1734	T6847	T6845	pobj	oviducts,from
R1735	T6848	T6847	prep	of,oviducts
R1736	T6849	T6850	amod	pregnant,females
R1737	T6850	T6848	pobj	females,of
R1738	T6851	T6842	prep	at,flushed
R1739	T6852	T6851	pobj	E2.5,at
R174	T854	T833	cc	and,deleted
R1740	T6853	T6852	cc	or,E2.5
R1741	T6854	T6852	conj	E3.5,E2.5
R1742	T6855	T6842	punct	", ",flushed
R1743	T6856	T6842	cc	and,flushed
R1744	T6857	T6842	conj	genotyped,flushed
R1745	T6858	T6857	prep	by,genotyped
R1746	T6859	T6858	pobj	means,by
R1747	T6860	T6859	prep	of,means
R1748	T6861	T6862	det	a,strategy
R1749	T6862	T6860	pobj	strategy,of
R175	T855	T833	conj	transmitted,deleted
R1750	T6863	T6862	amod	nested,strategy
R1751	T6864	T6862	compound	PCR,strategy
R1752	T6865	T6866	punct	(,Figure
R1753	T6866	T6857	parataxis	Figure,genotyped
R1754	T6867	T6866	nummod	4,Figure
R1755	T6868	T6866	punct	),Figure
R1756	T6869	T6842	punct	.,flushed
R1757	T6871	T6872	nsubj	Two,were
R1758	T6873	T6871	prep	of,Two
R1759	T6874	T6875	nummod	90,embryos
R176	T856	T857	det	this,allele
R1760	T6875	T6873	pobj	embryos,of
R1761	T6876	T6877	advmod	successfully,genotyped
R1762	T6877	T6875	amod	genotyped,embryos
R1763	T6878	T6875	amod	pre-implantation,embryos
R1764	T6879	T6872	attr	Capn2,were
R1765	T6880	T6879	punct	-,Capn2
R1766	T6881	T6879	punct	/,Capn2
R1767	T6882	T6879	punct	-,Capn2
R1768	T6883	T6872	punct	", ",were
R1769	T6884	T6885	punct	(,Figure
R177	T857	T855	dobj	allele,transmitted
R1770	T6885	T6872	parataxis	Figure,were
R1771	T6886	T6885	dep	Table,Figure
R1772	T6887	T6886	nummod	2,Table
R1773	T6888	T6885	punct	;,Figure
R1774	T6889	T6885	nummod	5,Figure
R1775	T6890	T6885	punct	),Figure
R1776	T6891	T6872	punct	.,were
R1777	T6893	T6894	nsubjpass	Both,isolated
R1778	T6895	T6893	prep	of,Both
R1779	T6896	T6897	det	these,embryos
R178	T858	T857	compound	mutant,allele
R1780	T6897	T6895	pobj	embryos,of
R1781	T6898	T6897	nmod	Capn2,embryos
R1782	T6899	T6898	punct	-,Capn2
R1783	T6900	T6898	punct	/,Capn2
R1784	T6901	T6898	punct	-,Capn2
R1785	T6902	T6894	auxpass	were,isolated
R1786	T6903	T6894	prep	at,isolated
R1787	T6904	T6905	det	the,stage
R1788	T6905	T6903	pobj	stage,at
R1789	T6906	T6907	nummod	8,cell
R179	T859	T855	prep	through,transmitted
R1790	T6907	T6905	compound	cell,stage
R1791	T6908	T6907	punct	-,cell
R1792	T6909	T6894	cc	and,isolated
R1793	T6910	T6911	aux	did,display
R1794	T6911	T6894	conj	display,isolated
R1795	T6912	T6911	neg	not,display
R1796	T6913	T6914	det	any,defects
R1797	T6914	T6911	dobj	defects,display
R1798	T6915	T6914	amod	obvious,defects
R1799	T6916	T6914	amod	morphological,defects
R18	T688	T687	punct	-,calpain
R180	T860	T861	det	the,germline
R1800	T6917	T6894	punct	.,isolated
R1801	T6919	T6920	nsubj	None,were
R1802	T6921	T6919	prep	of,None
R1803	T6922	T6923	det	the,embryos
R1804	T6923	T6921	pobj	embryos,of
R1805	T6924	T6923	nummod	46,embryos
R1806	T6925	T6926	advmod	successfully,genotyped
R1807	T6926	T6923	amod	genotyped,embryos
R1808	T6927	T6928	npadvmod	blastocyst,staged
R1809	T6928	T6923	amod	staged,embryos
R181	T861	T859	pobj	germline,through
R1810	T6929	T6928	punct	-,staged
R1811	T6930	T6920	attr	Capn2,were
R1812	T6931	T6930	punct	-,Capn2
R1813	T6932	T6930	punct	/,Capn2
R1814	T6933	T6930	punct	-,Capn2
R1815	T6934	T6920	punct	.,were
R1816	T6936	T6937	det	The,scarcity
R1817	T6937	T6938	nsubj	scarcity,suggested
R1818	T6939	T6937	prep	of,scarcity
R1819	T6940	T6941	npadvmod	Capn2,deficient
R182	T862	T861	compound	mouse,germline
R1820	T6941	T6943	amod	deficient,embryos
R1821	T6942	T6941	punct	-,deficient
R1822	T6943	T6939	pobj	embryos,of
R1823	T6944	T6943	acl	surviving,embryos
R1824	T6945	T6944	prep	to,surviving
R1825	T6946	T6947	det	the,stage
R1826	T6947	T6945	pobj	stage,to
R1827	T6948	T6949	nummod	8,cell
R1828	T6949	T6947	compound	cell,stage
R1829	T6950	T6949	punct	-,cell
R183	T863	T833	punct	.,deleted
R1830	T6951	T6952	mark	that,compromise
R1831	T6952	T6938	ccomp	compromise,suggested
R1832	T6953	T6954	det	the,loss
R1833	T6954	T6952	nsubj	loss,compromise
R1834	T6955	T6954	prep	of,loss
R1835	T6956	T6957	compound	m,calpain
R1836	T6957	T6959	compound	calpain,activity
R1837	T6958	T6957	punct	-,calpain
R1838	T6959	T6955	pobj	activity,of
R1839	T6960	T6952	aux	must,compromise
R184	T865	T866	nsubj	Breeding,failed
R1840	T6961	T6952	advmod	fatally,compromise
R1841	T6962	T6963	det	the,viability
R1842	T6963	T6952	dobj	viability,compromise
R1843	T6964	T6963	prep	of,viability
R1844	T6965	T6966	amod	early,embryos
R1845	T6966	T6964	pobj	embryos,of
R1846	T6967	T6938	punct	.,suggested
R1847	T6969	T6970	advmod	Furthermore,is
R1848	T6971	T6970	punct	", ",is
R1849	T6972	T6970	nsubj	it,is
R185	T867	T865	prep	of,Breeding
R1850	T6973	T6970	acomp	possible,is
R1851	T6974	T6975	mark	that,allowed
R1852	T6975	T6970	ccomp	allowed,is
R1853	T6976	T6975	nsubj	persistence,allowed
R1854	T6977	T6976	prep	of,persistence
R1855	T6978	T6979	det	some,transcript
R1856	T6979	T6977	pobj	transcript,of
R1857	T6980	T6981	advmod	maternally,derived
R1858	T6981	T6979	amod	derived,transcript
R1859	T6982	T6979	compound	mRNA,transcript
R186	T868	T869	amod	heterozygous,animals
R1860	T6983	T6979	cc	or,transcript
R1861	T6984	T6979	conj	protein,transcript
R1862	T6985	T6975	aux	might,allowed
R1863	T6986	T6975	aux	have,allowed
R1864	T6987	T6988	det	a,number
R1865	T6988	T6990	nsubj	number,survive
R1866	T6989	T6988	amod	small,number
R1867	T6990	T6975	ccomp	survive,allowed
R1868	T6991	T6988	prep	of,number
R1869	T6992	T6993	nmod	Capn2,embryos
R187	T869	T867	pobj	animals,of
R1870	T6993	T6991	pobj	embryos,of
R1871	T6994	T6992	punct	-,Capn2
R1872	T6995	T6992	punct	/,Capn2
R1873	T6996	T6992	punct	-,Capn2
R1874	T6997	T6990	aux	to,survive
R1875	T6998	T6990	prep	to,survive
R1876	T6999	T7000	det	the,stage
R1877	T7000	T6998	pobj	stage,to
R1878	T7001	T7000	compound	morula,stage
R1879	T7002	T7000	punct	-,stage
R188	T870	T871	aux	to,produce
R1880	T7003	T6970	punct	.,is
R1881	T7005	T7006	det	"Table 2 Genotype distribution of offspring derived from Capn2 transgenic mice. Cross (N) Age Genotype +/+ +/- -/- ND* (+/-)♀ × (+/-)♂ 199 3 weeks 23 176 0 0 (+/+)♀ × (+/-)♂ 122 3 weeks 50 72 0 0 (+/-)♀ × (+/+)♂ 103 3 weeks 28 75 0 0 (+/-)♀ × (+/-)♂ 9 E18.5 0 7 0 2 (+/-)♀ × (+/-)♂ 5 E17.5 0 5 0 0 (+/-)♀ × (+/-)♂ 9 E14.5 2 7 0 0 (+/-)♀ × (+/-)♂ 10 E11.5 3 7 0 0 (+/-)♀ × (+/-)♂ 7 E10.5 2 5 0 0 (+/-)♀ × (+/-)♂ 46 E3.5 7 39 0 0 (+/-)♀ × (+/-)♂ 48 E2.5 8 34 2 4 * ND, not determined Figure 4 Nested PCR strategy for genotyping of pre-implantation embryos. Due to the scarcity of extractable genetic material, a nested PCR strategy was developed in order to genotype pre-implantation embryos. Separate reactions were used to amplify a 429 bp fragment from the wild-type allele and a 389 bp segment from the mutant allele, both spanning the 3' end of the short (upstream) arm of the targeting vector. In both reactions, a 213 bp sequence located within the short arm was co-amplified with the 'diagnostic' products as an internal control. The same sense primers were used to amplify 'diagnostic' sequences in both reactions, whereas the antisense primers were allele-specific. The primers, represented by triangles, are depicted in two (nested) sets for each of the three reactions. Exons are represented by open, vertical rectangles, the PGK-Neo cassette by an open, horizontal rectangle, while thin vertical lines denote the boundaries of the short arm and the 5' end of the long (downstream) arm. A grey, horizontal rectangle delineates the segment of the wild-type allele that is replaced by the PGK-Neo cassette in the mutant allele. Figure 5 Genotyping of pre-implantation embryos. A nested PCR strategy was used to genotype embryos prior to implantation as described in Figure 4. Capn2+/- mice were mated and the date of fertilization established by the appearance of vaginal plugs. Blastocyst (E3.5) or 8-cell embryos (E2.5) were flushed from the oviducts and then digested with proteinase K. In separate reactions segments found exclusively in either the wild-type or mutant alleles were co-amplified with an internal control sequence, located in the short (upstream) arm of the targeting vector, which is found in both alleles. The final products were 429 bp for the wild-type allele, 389 bp for the mutant allele, and 213 bp for the internal control. (A) A representative example of the genotyping of blastocyst stage embryos. Embryos #1, 2, 4, 5, and 6 were Capn2+/- whereas embryos #3 and #7 were Capn2+/+, denoted by the absence of the 389 bp mutant signal. (B) An example of the genotyping of 8-cell embryos is shown. Embryos #1, 2, 3, 5, and 6 were Capn2+/- while embryo #4 was Capn2-/-, marked by the absence of the 429 bp wild-type signal. (M) denotes the molecular weight marker. The",genotyping
R1882	T7006	T7007	nsubjpass	genotyping,shown
R1883	T7008	T7006	nmod	results,genotyping
R1884	T7009	T7006	prep	for,genotyping
R1885	T7010	T7009	pobj	weanlings,for
R1886	T7011	T7010	punct	", ",weanlings
R1887	T7012	T7013	nmod	post-implantation,embryos
R1888	T7013	T7010	appos	embryos,weanlings
R1889	T7014	T7012	punct	", ",post-implantation
R189	T871	T866	xcomp	produce,failed
R1890	T7015	T7012	cc	and,post-implantation
R1891	T7016	T7012	conj	pre-implantation,post-implantation
R1892	T7017	T7007	auxpass	are,shown
R1893	T7018	T7007	prep	in,shown
R1894	T7019	T7018	pobj	Table,in
R1895	T7020	T7019	nummod	2,Table
R1896	T7021	T7007	punct	.,shown
R1897	T7023	T7024	advmod	Curiously,was
R1898	T7025	T7024	punct	", ",was
R1899	T7026	T7027	det	the,ratio
R19	T689	T690	advmod	ubiquitously,expressed
R190	T872	T873	amod	homozygous,offspring
R1900	T7027	T7024	nsubj	ratio,was
R1901	T7028	T7027	prep	of,ratio
R1902	T7029	T7028	pobj	Capn2,of
R1903	T7030	T7029	punct	+,Capn2
R1904	T7031	T7029	punct	/,Capn2
R1905	T7032	T7029	punct	+,Capn2
R1906	T7033	T7027	prep	to,ratio
R1907	T7034	T7035	nmod	Capn2,animals
R1908	T7035	T7033	pobj	animals,to
R1909	T7036	T7034	punct	+,Capn2
R191	T873	T871	dobj	offspring,produce
R1910	T7037	T7034	punct	/,Capn2
R1911	T7038	T7034	punct	-,Capn2
R1912	T7039	T7027	prep	from,ratio
R1913	T7040	T7041	amod	inbred,intercrosses
R1914	T7041	T7039	pobj	intercrosses,from
R1915	T7042	T7041	amod	heterozygous,intercrosses
R1916	T7043	T7044	advmod	substantially,less
R1917	T7044	T7024	acomp	less,was
R1918	T7045	T7044	prep	than,less
R1919	T7046	T7047	det	the,ratio
R192	T874	T873	nmod	mutant,offspring
R1920	T7047	T7045	pobj	ratio,than
R1921	T7048	T7047	amod	predicted,ratio
R1922	T7049	T7047	nummod	1,ratio
R1923	T7050	T7051	punct	:,2
R1924	T7051	T7049	prep	2,1
R1925	T7052	T7047	amod	Mendelian,ratio
R1926	T7053	T7024	punct	.,was
R1927	T7055	T7056	prep	In,observed
R1928	T7057	T7058	det	a,group
R1929	T7058	T7055	pobj	group,In
R193	T875	T873	amod	live,offspring
R1930	T7059	T7058	prep	of,group
R1931	T7060	T7061	nummod	199,animals
R1932	T7061	T7059	pobj	animals,of
R1933	T7062	T7061	acl	derived,animals
R1934	T7063	T7062	prep	from,derived
R1935	T7064	T7065	compound	heterozygote,breeding
R1936	T7065	T7063	pobj	breeding,from
R1937	T7066	T7067	punct	(,litters
R1938	T7067	T7065	parataxis	litters,breeding
R1939	T7068	T7067	nummod	33,litters
R194	T876	T873	cc	or,offspring
R1940	T7069	T7067	punct	),litters
R1941	T7070	T7056	punct	", ",observed
R1942	T7071	T7072	nummod	23,Capn2
R1943	T7072	T7056	nsubjpass	Capn2,observed
R1944	T7073	T7072	punct	+,Capn2
R1945	T7074	T7072	punct	/,Capn2
R1946	T7075	T7072	punct	+,Capn2
R1947	T7076	T7077	punct	(,%
R1948	T7077	T7072	parataxis	%,Capn2
R1949	T7078	T7077	nummod	11.6,%
R195	T877	T878	amod	implanted,embryos
R1950	T7079	T7077	punct	),%
R1951	T7080	T7072	cc	and,Capn2
R1952	T7081	T7082	nummod	176,Capn2
R1953	T7082	T7087	nmod	Capn2,animals
R1954	T7083	T7084	punct	(,%
R1955	T7084	T7082	parataxis	%,Capn2
R1956	T7085	T7084	nummod	88.4,%
R1957	T7086	T7084	punct	),%
R1958	T7087	T7072	conj	animals,Capn2
R1959	T7088	T7082	punct	+,Capn2
R196	T878	T873	conj	embryos,offspring
R1960	T7089	T7082	punct	/,Capn2
R1961	T7090	T7082	punct	-,Capn2
R1962	T7091	T7056	auxpass	were,observed
R1963	T7092	T7056	punct	.,observed
R1964	T7094	T7095	det	The,ratio
R1965	T7095	T7096	nsubj	ratio,was
R1966	T7097	T7095	prep	of,ratio
R1967	T7098	T7097	pobj	Capn2,of
R1968	T7099	T7098	punct	+,Capn2
R1969	T7100	T7098	punct	/,Capn2
R197	T879	T866	punct	.,failed
R1970	T7101	T7098	punct	+,Capn2
R1971	T7102	T7095	prep	to,ratio
R1972	T7103	T7104	nmod	Capn2,animals
R1973	T7104	T7102	pobj	animals,to
R1974	T7105	T7103	punct	+,Capn2
R1975	T7106	T7103	punct	/,Capn2
R1976	T7107	T7103	punct	-,Capn2
R1977	T7108	T7095	prep	among,ratio
R1978	T7109	T7108	pobj	males,among
R1979	T7110	T7111	punct	(,%
R198	T881	T882	det	A,protocol
R1980	T7111	T7109	parataxis	%,males
R1981	T7112	T7111	nummod	14,%
R1982	T7113	T7111	prep	to,%
R1983	T7114	T7115	nummod	82,%
R1984	T7115	T7113	pobj	%,to
R1985	T7116	T7111	punct	),%
R1986	T7117	T7109	cc	or,males
R1987	T7118	T7109	conj	females,males
R1988	T7202	T7183	advcl	was,reported
R1989	T7119	T7120	punct	(,%
R199	T882	T886	nsubjpass	protocol,established
R1990	T7203	T7204	det	the,skewing
R1991	T7120	T7118	parataxis	%,females
R1992	T7204	T7202	nsubj	skewing,was
R1993	T7121	T7120	nummod	13,%
R1994	T7205	T7204	compound	genotype,skewing
R1995	T7206	T7202	neg	not,was
R1996	T7207	T7202	prep	as,was
R1997	T7122	T7120	prep	to,%
R1998	T7208	T7207	amod	extreme,as
R1999	T7209	T7210	punct	[,32
R20	T690	T691	amod	expressed,proteases
R200	T883	T882	amod	nested,protocol
R2000	T7123	T7124	nummod	90,%
R2001	T7210	T7202	parataxis	32,was
R2002	T7211	T7210	punct	],32
R2003	T7212	T7183	punct	.,reported
R2004	T7124	T7122	pobj	%,to
R2005	T7214	T7215	nsubj	Crosses,resulted
R2006	T7125	T7120	punct	),%
R2007	T7216	T7214	prep	between,Crosses
R2008	T7217	T7218	nmod	Capn2,animals
R2009	T7218	T7216	pobj	animals,between
R201	T884	T882	compound	PCR,protocol
R2010	T7219	T7217	punct	+,Capn2
R2011	T7220	T7217	punct	/,Capn2
R2012	T7126	T7096	advmod	essentially,was
R2013	T7221	T7217	punct	+,Capn2
R2014	T7222	T7217	cc	and,Capn2
R2015	T7223	T7217	conj	Capn2,Capn2
R2016	T7224	T7223	punct	+,Capn2
R2017	T7127	T7128	advmod	the,same
R2018	T7128	T7096	acomp	same,was
R2019	T7129	T7130	mark	is,was
R202	T885	T882	compound	genotyping,protocol
R2020	T7225	T7223	punct	/,Capn2
R2021	T7130	T7128	advcl	was,same
R2022	T7226	T7223	punct	-,Capn2
R2023	T7227	T7215	advmod	also,resulted
R2024	T7228	T7215	prep	in,resulted
R2025	T7131	T7130	nsubj	it,was
R2026	T7229	T7230	det	a,proportion
R2027	T7230	T7228	pobj	proportion,in
R2028	T7132	T7130	prep	for,was
R2029	T7231	T7230	amod	greater,proportion
R203	T887	T886	auxpass	was,established
R2030	T7232	T7233	mark	than,expected
R2031	T7233	T7231	advcl	expected,greater
R2032	T7133	T7134	det	the,population
R2033	T7234	T7230	prep	of,proportion
R2034	T7235	T7236	nmod	Capn2,animals
R2035	T7134	T7132	pobj	population,for
R2036	T7236	T7234	pobj	animals,of
R2037	T7237	T7235	punct	+,Capn2
R2038	T7238	T7235	punct	/,Capn2
R2039	T7135	T7134	amod	combined,population
R204	T888	T886	punct	", ",established
R2040	T7239	T7235	punct	-,Capn2
R2041	T7240	T7241	punct	(,Table
R2042	T7241	T7215	parataxis	Table,resulted
R2043	T7136	T7096	punct	", ",was
R2044	T7242	T7241	nummod	2,Table
R2045	T7243	T7241	punct	),Table
R2046	T7244	T7215	punct	.,resulted
R2047	T7137	T7096	cc	and,was
R2048	T7246	T7247	det	An,degree
R2049	T7247	T7250	nsubjpass	degree,seen
R205	T889	T886	cc	and,established
R2050	T7138	T7139	expl	there,were
R2051	T7248	T7249	advmod	even,higher
R2052	T7249	T7247	amod	higher,degree
R2053	T7139	T7096	conj	were,was
R2054	T7251	T7247	prep	of,degree
R2055	T7252	T7253	nmod	Capn2,over-representation
R2056	T7140	T7141	det	an,average
R2057	T7253	T7251	pobj	over-representation,of
R2058	T7254	T7252	punct	+,Capn2
R2059	T7255	T7252	punct	/,Capn2
R206	T890	T891	amod	homozygous,embryos
R2060	T7141	T7139	attr	average,were
R2061	T7256	T7252	punct	-,Capn2
R2062	T7257	T7250	auxpass	was,seen
R2063	T7258	T7259	advmod	when,came
R2064	T7259	T7250	advcl	came,seen
R2065	T7260	T7261	det	the,allele
R2066	T7261	T7259	nsubj	allele,came
R2067	T7142	T7141	prep	of,average
R2068	T7262	T7261	compound	mutant,allele
R2069	T7263	T7264	det	the,mother
R207	T891	T894	nsubjpass	embryos,detected
R2070	T7264	T7259	dobj	mother,came
R2071	T7143	T7144	nummod	six,pups
R2072	T7265	T7266	punct	(,%
R2073	T7266	T7259	parataxis	%,came
R2074	T7144	T7142	pobj	pups,of
R2075	T7267	T7266	nummod	73,%
R2076	T7268	T7266	punct	),%
R2077	T7269	T7250	prep	compared,seen
R2078	T7145	T7144	prep	per,pups
R2079	T7270	T7269	prep	to,compared
R208	T892	T891	compound	preimplantation,embryos
R2080	T7271	T7272	advmod	when,came
R2081	T7146	T7145	pobj	litter,per
R2082	T7272	T7270	pcomp	came,to
R2083	T7273	T7272	nsubj	it,came
R2084	T7147	T7141	punct	", ",average
R2085	T7148	T7149	dep	which,is
R2086	T7274	T7272	prep	from,came
R2087	T7149	T7141	relcl	is,average
R2088	T7275	T7276	det	the,father
R2089	T7276	T7274	pobj	father,from
R209	T893	T891	compound	mutant,embryos
R2090	T7150	T7149	acomp	normal,is
R2091	T7277	T7278	punct	(,%
R2092	T7278	T7272	parataxis	%,came
R2093	T7279	T7278	nummod	59,%
R2094	T7151	T7149	prep	for,is
R2095	T7280	T7278	punct	),%
R2096	T7281	T7250	punct	.,seen
R2097	T7152	T7153	det	this,strain
R2098	T7283	T7284	prep	In,compared
R2099	T7153	T7151	pobj	strain,for
R21	T691	T684	attr	proteases,are
R210	T894	T886	conj	detected,established
R2100	T7285	T7286	det	these,crosses
R2101	T7286	T7283	pobj	crosses,In
R2102	T7154	T7153	compound	background,strain
R2103	T7287	T7288	det	the,ratios
R2104	T7288	T7284	nsubj	ratios,compared
R2105	T7289	T7288	prep	of,ratios
R2106	T7155	T7139	punct	.,were
R2107	T7290	T7289	pobj	Capn2,of
R2108	T7291	T7290	punct	+,Capn2
R2109	T7157	T7158	det	A,proportion
R211	T895	T894	auxpass	were,detected
R2110	T7292	T7290	punct	/,Capn2
R2111	T7293	T7290	punct	+,Capn2
R2112	T7294	T7288	prep	to,ratios
R2113	T7295	T7296	nmod	Capn2,animals
R2114	T7296	T7294	pobj	animals,to
R2115	T7297	T7295	punct	+,Capn2
R2116	T7298	T7295	punct	/,Capn2
R2117	T7158	T7162	nsubj	proportion,was
R2118	T7299	T7295	punct	-,Capn2
R2119	T7300	T7288	prep	among,ratios
R212	T896	T894	prep	at,detected
R2120	T7301	T7300	pobj	males,among
R2121	T7159	T7158	amod	larger,proportion
R2122	T7302	T7301	cc	or,males
R2123	T7303	T7301	conj	females,males
R2124	T7304	T7284	advmod	well,compared
R2125	T7160	T7161	mark	than,expected
R2126	T7305	T7284	prep	with,compared
R2127	T7306	T7307	det	the,ratio
R2128	T7161	T7159	advcl	expected,larger
R2129	T7163	T7164	nmod	Capn2,animals
R213	T897	T898	det	the,morula
R2130	T7164	T7158	npadvmod	animals,proportion
R2131	T7307	T7305	pobj	ratio,with
R2132	T7165	T7163	punct	+,Capn2
R2133	T7308	T7284	prep	in,compared
R2134	T7309	T7310	det	the,populations
R2135	T7310	T7308	pobj	populations,in
R2136	T7166	T7163	punct	/,Capn2
R2137	T7311	T7310	amod	combined,populations
R2138	T7312	T7284	punct	;,compared
R2139	T7167	T7163	punct	-,Capn2
R214	T898	T896	pobj	morula,at
R2140	T7313	T7314	nummod	77,%
R2141	T7314	T7315	nsubj	%,were
R2142	T7315	T7284	conj	were,compared
R2143	T7168	T7162	advmod	also,was
R2144	T7316	T7314	prep	of,%
R2145	T7317	T7316	pobj	males,of
R2146	T7318	T7314	cc	and,%
R2147	T7169	T7162	acomp	evident,was
R2148	T7319	T7320	nummod	69,%
R2149	T7320	T7314	conj	%,%
R215	T899	T894	cc	but,detected
R2150	T7170	T7162	prep	in,was
R2151	T7321	T7320	prep	of,%
R2152	T7322	T7321	pobj	females,of
R2153	T7323	T7315	attr	Capn2,were
R2154	T7171	T7172	preconj	both,embryos
R2155	T7324	T7323	punct	+,Capn2
R2156	T7325	T7323	punct	/,Capn2
R2157	T7326	T7323	punct	-,Capn2
R2158	T7172	T7170	pobj	embryos,in
R2159	T7327	T7328	advmod	when,came
R216	T900	T894	conj	not,detected
R2160	T7328	T7315	advcl	came,were
R2161	T7329	T7330	det	the,allele
R2162	T7173	T7172	nmod	post-implantation,embryos
R2163	T7330	T7328	nsubj	allele,came
R2164	T7331	T7330	compound	mutant,allele
R2165	T7332	T7328	prep	from,came
R2166	T7333	T7334	det	the,mother
R2167	T7334	T7332	pobj	mother,from
R2168	T7335	T7315	punct	", ",were
R2169	T7174	T7173	cc	and,post-implantation
R217	T901	T900	prep	at,not
R2170	T7336	T7315	cc	and,were
R2171	T7337	T7338	nummod	62,%
R2172	T7338	T7339	nsubj	%,were
R2173	T7175	T7173	conj	pre-implantation,post-implantation
R2174	T7339	T7315	conj	were,were
R2175	T7340	T7338	prep	of,%
R2176	T7341	T7340	pobj	males,of
R2177	T7176	T7177	punct	(,Table
R2178	T7342	T7338	cc	and,%
R2179	T7343	T7344	nummod	55,%
R218	T902	T903	det	the,blastocyts
R2180	T7344	T7338	conj	%,%
R2181	T7177	T7162	parataxis	Table,was
R2182	T7345	T7344	prep	of,%
R2183	T7346	T7345	pobj	females,of
R2184	T7347	T7339	attr	Capn2,were
R2185	T7178	T7177	nummod	2,Table
R2186	T7348	T7347	punct	+,Capn2
R2187	T7349	T7347	punct	/,Capn2
R2188	T7350	T7347	punct	-,Capn2
R2189	T7179	T7177	punct	),Table
R219	T903	T901	pobj	blastocyts,at
R2190	T7351	T7352	advmod	when,came
R2191	T7352	T7339	advcl	came,were
R2192	T7180	T7162	punct	.,was
R2193	T7353	T7354	det	the,allele
R2194	T7354	T7352	nsubj	allele,came
R2195	T7355	T7354	compound	mutant,allele
R2196	T7182	T7183	advmod	Interestingly,reported
R2197	T7356	T7352	prep	from,came
R2198	T7184	T7183	punct	", ",reported
R2199	T7357	T7358	det	the,father
R22	T692	T691	acl	implicated,proteases
R220	T904	T900	dobj	stage,not
R2200	T7358	T7356	pobj	father,from
R2201	T7185	T7186	det	a,over-representation
R2202	T7359	T7339	punct	.,were
R2203	T7186	T7183	nsubjpass	over-representation,reported
R2204	T7187	T7186	amod	similar,over-representation
R2205	T7188	T7186	prep	of,over-representation
R2206	T7189	T7190	amod	heterozygous,animals
R2207	T7190	T7188	pobj	animals,of
R2208	T7191	T7183	auxpass	was,reported
R2209	T7192	T7183	advmod	also,reported
R221	T905	T894	punct	.,detected
R2210	T7193	T7183	prep	in,reported
R2211	T7194	T7193	pobj	one,in
R2212	T7195	T7194	prep	of,one
R2213	T7196	T7197	det	the,lines
R2214	T7197	T7195	pobj	lines,of
R2215	T7198	T7197	nmod	Capn4,lines
R2216	T7199	T7197	amod	transgenic,lines
R2217	T7200	T7183	punct	", ",reported
R2218	T7201	T7202	mark	though,was
R222	T909	T910	nsubj	We,conclude
R2221	T7704	T7705	amod	Attempted,generation
R2222	T7706	T7705	prep	of,generation
R2223	T7707	T7708	nmod	Capn2,cells
R2224	T7708	T7706	pobj	cells,of
R2225	T7709	T7707	punct	-,Capn2
R2226	T7710	T7707	punct	/,Capn2
R2227	T7711	T7707	punct	-,Capn2
R2228	T7712	T7708	compound	ES,cells
R2229	T7714	T7715	nmod	Capn2,cells
R223	T911	T912	mark	that,results
R2230	T7715	T7720	nsubjpass	cells,subjected
R2231	T7716	T7714	punct	+,Capn2
R2232	T7717	T7714	punct	/,Capn2
R2233	T7718	T7714	punct	-,Capn2
R2234	T7719	T7715	compound	ES,cells
R2235	T7721	T7720	auxpass	were,subjected
R2236	T7722	T7720	prep	to,subjected
R2237	T7723	T7724	amod	clonal,selection
R2238	T7724	T7722	pobj	selection,to
R2239	T7725	T7720	prep	in,subjected
R224	T912	T910	ccomp	results,conclude
R2240	T7726	T7727	det	the,presence
R2241	T7727	T7725	pobj	presence,in
R2242	T7728	T7727	prep	of,presence
R2243	T7729	T7730	nummod	2,mg
R2244	T7730	T7731	nmod	mg,G418
R2245	T7731	T7728	pobj	G418,of
R2246	T7732	T7733	punct	/,mL
R2247	T7733	T7730	prep	mL,mg
R2248	T7734	T7727	prep	in,presence
R2249	T7735	T7734	pobj	attempts,in
R225	T913	T914	amod	homozygous,disruption
R2250	T7736	T7737	aux	to,generate
R2251	T7737	T7735	acl	generate,attempts
R2252	T7738	T7739	amod	homozygous,cells
R2253	T7739	T7737	dobj	cells,generate
R2254	T7740	T7739	compound	mutant,cells
R2255	T7741	T7737	prep	by,generate
R2256	T7742	T7743	compound	gene,conversion
R2257	T7743	T7741	pobj	conversion,by
R2258	T7744	T7720	punct	.,subjected
R2259	T7746	T7747	det	This,procedure
R226	T914	T912	nsubj	disruption,results
R2260	T7747	T7748	nsubjpass	procedure,applied
R2261	T7749	T7748	aux	has,applied
R2262	T7750	T7748	auxpass	been,applied
R2263	T7751	T7748	advmod	extensively,applied
R2264	T7752	T7748	prep	to,applied
R2265	T7753	T7754	amod	targeted,cells
R2266	T7754	T7752	pobj	cells,to
R2267	T7755	T7754	compound	ES,cells
R2268	T7756	T7748	cc	and,applied
R2269	T7757	T7758	auxpass	was,achieved
R227	T915	T914	prep	of,disruption
R2270	T7758	T7748	conj	achieved,applied
R2271	T7759	T7758	advmod	readily,achieved
R2272	T7760	T7758	prep	in,achieved
R2273	T7761	T7762	det	the,case
R2274	T7762	T7760	pobj	case,in
R2275	T7763	T7762	prep	of,case
R2276	T7764	T7765	nmod	Capn4,cells
R2277	T7765	T7763	pobj	cells,of
R2278	T7766	T7764	punct	+,Capn4
R2279	T7767	T7764	punct	/,Capn4
R228	T916	T917	det	the,gene
R2280	T7768	T7764	punct	-,Capn4
R2281	T7769	T7765	compound	ES,cells
R2282	T7770	T7771	punct	[,31
R2283	T7771	T7758	parataxis	31,achieved
R2284	T7772	T7771	punct	],31
R2285	T7773	T7748	punct	.,applied
R2286	T7775	T7776	prep	In,isolated
R2287	T7777	T7778	det	this,case
R2288	T7778	T7775	pobj	case,In
R2289	T7779	T7776	punct	", ",isolated
R229	T917	T915	pobj	gene,of
R2290	T7780	T7776	advmod	however,isolated
R2291	T7781	T7776	punct	", ",isolated
R2292	T7782	T7783	det	no,cells
R2293	T7783	T7776	nsubjpass	cells,isolated
R2294	T7784	T7783	nmod	Capn2,cells
R2295	T7785	T7784	punct	-,Capn2
R2296	T7786	T7784	punct	/,Capn2
R2297	T7787	T7784	punct	-,Capn2
R2298	T7788	T7783	compound	ES,cells
R2299	T7789	T7776	auxpass	were,isolated
R23	T693	T692	prep	in,implicated
R230	T918	T917	compound	Capn2,gene
R2300	T7790	T7776	prep	in,isolated
R2301	T7791	T7790	pobj	screens,in
R2302	T7792	T7791	prep	of,screens
R2303	T7793	T7794	nummod	126,clones
R2304	T7794	T7792	pobj	clones,of
R2305	T7795	T7796	npadvmod	drug,resistant
R2306	T7796	T7794	amod	resistant,clones
R2307	T7797	T7796	punct	-,resistant
R2308	T7798	T7776	punct	.,isolated
R2309	T7800	T7801	det	The,inability
R231	T919	T912	prep	in,results
R2310	T7801	T7802	nsubj	inability,suggested
R2311	T7803	T7804	aux	to,isolate
R2312	T7804	T7801	acl	isolate,inability
R2313	T7805	T7806	nmod	Capn2,cells
R2314	T7806	T7804	dobj	cells,isolate
R2315	T7807	T7805	punct	-,Capn2
R2316	T7808	T7805	punct	/,Capn2
R2317	T7809	T7805	punct	-,Capn2
R2318	T7810	T7806	compound	ES,cells
R2319	T7811	T7804	punct	", ",isolate
R232	T920	T921	amod	pre-implantation,lethality
R2320	T7812	T7804	prep	in,isolate
R2321	T7813	T7812	pobj	concert,in
R2322	T7814	T7813	prep	with,concert
R2323	T7815	T7816	det	the,absence
R2324	T7816	T7814	pobj	absence,with
R2325	T7817	T7816	prep	of,absence
R2326	T7818	T7819	nmod	Capn2,embryos
R2327	T7819	T7817	pobj	embryos,of
R2328	T7820	T7818	punct	-,Capn2
R2329	T7821	T7818	punct	/,Capn2
R233	T921	T919	pobj	lethality,in
R2330	T7822	T7818	punct	-,Capn2
R2331	T7823	T7816	prep	beyond,absence
R2332	T7824	T7825	det	the,stage
R2333	T7825	T7823	pobj	stage,beyond
R2334	T7826	T7827	nummod	8,cell
R2335	T7827	T7825	compound	cell,stage
R2336	T7828	T7827	punct	-,cell
R2337	T7829	T7802	punct	", ",suggested
R2338	T7830	T7831	mark	that,be
R2339	T7831	T7802	ccomp	be,suggested
R234	T922	T921	amod	embryonic,lethality
R2340	T7832	T7833	compound	m,calpain
R2341	T7833	T7835	compound	calpain,activity
R2342	T7834	T7833	punct	-,calpain
R2343	T7835	T7831	nsubj	activity,be
R2344	T7836	T7831	aux	might,be
R2345	T7837	T7831	acomp	essential,be
R2346	T7838	T7837	prep	for,essential
R2347	T7839	T7840	compound	cell,viability
R2348	T7840	T7838	pobj	viability,for
R2349	T7841	T7840	cc	or,viability
R235	T923	T912	prep	between,results
R2350	T7842	T7843	det	the,establishment
R2351	T7843	T7840	conj	establishment,viability
R2352	T7844	T7843	prep	of,establishment
R2353	T7845	T7846	amod	viable,clones
R2354	T7846	T7844	pobj	clones,of
R2355	T7847	T7846	compound	ES,clones
R2356	T7848	T7846	compound	cell,clones
R2357	T7849	T7802	punct	.,suggested
R236	T924	T925	det	the,stage
R237	T925	T923	pobj	stage,between
R2370	T12875	T12876	det	the,story
R2373	T12816	T12817	mark	Although,identified
R2374	T12876	T12870	nsubjpass	story,complicated
R2375	T12877	T12870	aux	has,complicated
R2376	T12817	T12822	advcl	identified,remained
R2377	T12818	T12819	compound	calpain,activity
R2378	T12878	T12870	auxpass	been,complicated
R2379	T12819	T12817	nsubjpass	activity,identified
R238	T926	T925	nmod	morula,stage
R2380	T12820	T12817	auxpass	was,identified
R2381	T12821	T12817	advmod	first,identified
R2382	T12879	T12870	advmod	further,complicated
R2383	T12823	T12824	nummod	four,decades
R2384	T12880	T12870	agent	by,complicated
R2385	T12824	T12825	npadvmod	decades,ago
R2386	T12825	T12817	advmod	ago,identified
R2387	T12826	T12822	punct	", ",remained
R2388	T12881	T12882	det	the,discovery
R2389	T12827	T12828	det	a,understanding
R239	T927	T926	cc	and,morula
R2390	T12828	T12822	nsubj	understanding,remained
R2391	T12882	T12880	pobj	discovery,by
R2392	T12829	T12828	amod	clear,understanding
R2393	T12830	T12828	prep	of,understanding
R2394	T12831	T12832	det	the,substrates
R2395	T12883	T12882	prep	of,discovery
R2396	T12884	T12885	det	a,number
R2397	T12832	T12830	pobj	substrates,of
R2398	T12885	T12883	pobj	number,of
R2399	T12833	T12832	cc	and,substrates
R24	T694	T695	amod	cellular,migration
R240	T928	T926	conj	blastocyst,morula
R2400	T12834	T12832	conj	functions,substrates
R2401	T12886	T12885	prep	of,number
R2402	T12835	T12832	prep	of,substrates
R2403	T12836	T12837	det	the,enzymes
R2404	T12887	T12888	advmod	previously,unknown
R2405	T12837	T12835	pobj	enzymes,of
R2406	T12838	T12822	aux	has,remained
R2407	T12839	T12822	acomp	elusive,remained
R2408	T12840	T12822	punct	.,remained
R2409	T12842	T12843	prep	In,been
R241	T929	T910	punct	.,conclude
R2410	T12844	T12845	amod	large,part
R2411	T12845	T12842	pobj	part,In
R2412	T12846	T12843	punct	", ",been
R2413	T12847	T12843	nsubj	this,been
R2414	T12848	T12843	aux	has,been
R2415	T12849	T12843	prep	due,been
R2416	T12850	T12849	pcomp	to,due
R2417	T12888	T12889	amod	unknown,isoforms
R2418	T12851	T12852	det	the,lack
R2419	T12852	T12849	pobj	lack,due
R242	T931	T932	nsubj	This,establishes
R2420	T12853	T12852	prep	of,lack
R2421	T12854	T12853	pobj	inhibitors,of
R2422	T12855	T12854	amod	capable,inhibitors
R2423	T12856	T12855	prep	of,capable
R2424	T12889	T12886	pobj	isoforms,of
R2425	T12857	T12858	advmod	specifically,regulating
R2426	T12858	T12856	pcomp	regulating,of
R2427	T12859	T12858	advmod	down,regulating
R2428	T12890	T12891	dep	which,regulated
R2429	T12860	T12858	punct	-,regulating
R243	T933	T934	mark	that,have
R2430	T12861	T12862	det	the,calpains
R2431	T12862	T12858	dobj	calpains,regulating
R2432	T12891	T12889	relcl	regulated,isoforms
R2433	T12863	T12858	prep	without,regulating
R2434	T12864	T12863	pcomp	affecting,without
R2435	T12865	T12866	amod	other,proteases
R2436	T12892	T12891	aux	may,regulated
R2437	T12866	T12864	dobj	proteases,affecting
R2438	T12867	T12843	punct	.,been
R2439	T12893	T12891	auxpass	be,regulated
R244	T934	T932	advcl	have,establishes
R2440	T12869	T12870	prep	In,complicated
R2441	T12894	T12891	advmod	differently,regulated
R2442	T12871	T12872	det	the,decade
R2443	T12872	T12869	pobj	decade,In
R2444	T12873	T12872	amod	past,decade
R2445	T12895	T12891	cc	and,regulated
R2446	T12874	T12870	punct	", ",complicated
R2447	T12896	T12891	conj	have,regulated
R2448	T12897	T12898	amod	different,specificity
R2449	T12898	T12896	dobj	specificity,have
R245	T935	T936	compound	μ,calpain
R2450	T12899	T12898	compound	substrate,specificity
R2451	T12981	T12979	nsubjpass	subunit,required
R2452	T12900	T12870	punct	.,complicated
R2453	T12982	T12981	amod	small,subunit
R2454	T12983	T12979	auxpass	is,required
R2455	T12902	T12903	compound	Gene,targeting
R2456	T12984	T12979	prep	for,required
R2457	T12985	T12986	preconj	both,calpain
R2458	T12903	T12904	nsubj	targeting,provided
R2459	T12905	T12903	prep	in,targeting
R246	T936	T934	nsubj	calpain,have
R2460	T12986	T12984	pobj	calpain,for
R2461	T12987	T12986	nmod	μ,calpain
R2462	T12988	T12987	punct	-,μ
R2463	T12989	T12987	cc	and,μ
R2464	T12990	T12987	conj	m,μ
R2465	T12991	T12986	punct	-,calpain
R2466	T12992	T12974	punct	", ",supported
R2467	T12993	T12974	cc	and,supported
R2468	T12994	T12995	advmod	furthermore,suggested
R2469	T12906	T12905	pobj	mice,in
R247	T937	T936	punct	-,calpain
R2470	T12995	T12974	conj	suggested,supported
R2471	T12996	T12997	nummod	four,possibilities
R2472	T12997	T12995	dobj	possibilities,suggested
R2473	T12907	T12904	aux	has,provided
R2474	T12998	T12997	prep	regarding,possibilities
R2475	T12999	T13000	poss	their,requirement
R2476	T13000	T12998	pobj	requirement,regarding
R2477	T12908	T12909	det	a,approach
R2478	T13001	T13000	prep	for,requirement
R2479	T13002	T13003	amod	embryonic,development
R248	T938	T936	cc	and,calpain
R2480	T13003	T13001	pobj	development,for
R2481	T12909	T12904	dobj	approach,provided
R2482	T13004	T12975	punct	: ,required
R2483	T13005	T12975	meta	1,required
R2484	T13006	T13005	punct	),1
R2485	T12910	T12909	amod	powerful,approach
R2486	T13007	T13008	det	both,isoforms
R2487	T13008	T12975	nsubjpass	isoforms,required
R2488	T12911	T12912	aux	to,examine
R2489	T13009	T12975	auxpass	were,required
R249	T939	T940	compound	m,calpain
R2490	T13010	T12975	punct	;,required
R2491	T13011	T13012	meta	2,required
R2492	T13012	T12975	conj	required,required
R2493	T13013	T13011	punct	),2
R2494	T12912	T12909	advcl	examine,approach
R2495	T13014	T13015	compound	μ,calpain
R2496	T13015	T13012	nsubjpass	calpain,required
R2497	T13016	T13015	punct	-,calpain
R2498	T12913	T12914	det	the,roles
R2499	T13017	T13012	auxpass	was,required
R25	T695	T693	pobj	migration,in
R250	T940	T936	conj	calpain,calpain
R2500	T13018	T13012	punct	;,required
R2501	T13019	T13020	meta	3,required
R2502	T12914	T12912	dobj	roles,examine
R2503	T13020	T13012	conj	required,required
R2504	T13021	T13019	punct	),3
R2505	T13022	T13023	compound	m,calpain
R2506	T12915	T12914	amod	physiologic,roles
R2507	T13023	T13020	nsubjpass	calpain,required
R2508	T13024	T13023	punct	-,calpain
R2509	T13025	T13020	auxpass	was,required
R251	T941	T940	punct	-,calpain
R2510	T12916	T12914	prep	of,roles
R2511	T13026	T13020	punct	", ",required
R2512	T13027	T13020	cc	or,required
R2513	T13028	T13029	meta	4,are
R2514	T13029	T13020	conj	are,required
R2515	T13030	T13028	punct	),4
R2516	T13031	T13032	nmod	μ,calpain
R2517	T13032	T13029	nsubj	calpain,are
R2518	T13033	T13031	punct	-,μ
R2519	T12917	T12918	amod	individual,isoforms
R252	T942	T943	amod	distinct,functions
R2520	T13034	T13031	cc	and,μ
R2521	T13035	T13031	conj	m,μ
R2522	T13036	T13032	punct	-,calpain
R2523	T12918	T12916	pobj	isoforms,of
R2524	T13037	T13029	acomp	redundant,are
R2525	T13038	T13029	punct	", ",are
R2526	T13039	T13029	cc	and,are
R2527	T12919	T12918	compound	calpain,isoforms
R2528	T13040	T13041	nsubjpass	one,required
R2529	T13041	T13029	conj	required,are
R253	T943	T934	dobj	functions,have
R2530	T13042	T13040	cc	or,one
R2531	T12920	T12904	punct	.,provided
R2532	T13043	T13044	det	the,other
R2533	T13044	T13045	nmod	other,isoform
R2534	T13045	T13040	conj	isoform,one
R2535	T12922	T12923	nsubjpass	This,used
R2536	T13046	T13041	auxpass	was,required
R2537	T13047	T12975	punct	.,required
R2538	T13049	T13050	det	These,options
R2539	T12924	T12923	auxpass	was,used
R254	T944	T934	punct	", ",have
R2540	T13050	T13051	nsubjpass	options,narrowed
R2541	T13052	T13051	auxpass	were,narrowed
R2542	T13053	T13051	prt	down,narrowed
R2543	T13054	T13051	agent	by,narrowed
R2544	T12925	T12923	advmod	first,used
R2545	T13055	T13056	det	the,observation
R2546	T13056	T13054	pobj	observation,by
R2547	T13057	T13056	amod	subsequent,observation
R2548	T13058	T13059	mark	that,were
R2549	T12926	T12927	aux	to,disrupt
R255	T945	T934	cc	and,have
R2550	T12927	T12923	advcl	disrupt,used
R2551	T13059	T13056	acl	were,observation
R2552	T12928	T12929	det	the,gene
R2553	T13060	T13061	nmod	Capn1,mice
R2554	T13061	T13059	nsubj	mice,were
R2555	T13062	T13060	punct	-,Capn1
R2556	T13063	T13060	punct	/,Capn1
R2557	T12929	T12927	dobj	gene,disrupt
R2558	T13064	T13060	punct	-,Capn1
R2559	T13065	T13061	punct	", ",mice
R256	T946	T947	mark	that,is
R2560	T12930	T12929	compound	Capn4,gene
R2561	T13066	T13067	dep	which,lack
R2562	T13067	T13061	relcl	lack,mice
R2563	T13068	T13069	det	the,subunit
R2564	T13069	T13067	dobj	subunit,lack
R2565	T13070	T13071	nmod	μ,calpain
R2566	T13071	T13069	nmod	calpain,subunit
R2567	T13072	T13071	punct	-,calpain
R2568	T13073	T13069	amod	catalytic,subunit
R2569	T13074	T13059	punct	", ",were
R257	T947	T934	conj	is,have
R2570	T13075	T13059	acomp	healthy,were
R2571	T13076	T13075	cc	and,healthy
R2572	T12931	T12923	punct	", ",used
R2573	T13077	T13075	conj	fertile,healthy
R2574	T13078	T13059	punct	", ",were
R2575	T13079	T13080	mark	although,observed
R2576	T12932	T12923	advcl	encoding,used
R2577	T13080	T13059	advcl	observed,were
R2578	T13081	T13082	nmod	platelet,aggregation
R2579	T12933	T12934	det	the,subunit
R258	T948	T949	compound	m,calpain
R2580	T13082	T13083	nmod	aggregation,defects
R2581	T13083	T13080	nsubjpass	defects,observed
R2582	T13084	T13082	cc	and,aggregation
R2583	T13085	T13086	compound	clot,retraction
R2584	T12934	T12932	dobj	subunit,encoding
R2585	T13086	T13082	conj	retraction,aggregation
R2586	T12935	T12934	amod	small,subunit
R2587	T12936	T12934	amod	regulatory,subunit
R2588	T12937	T12934	amod	common,subunit
R2589	T13087	T13080	auxpass	were,observed
R259	T949	T947	nsubj	calpain,is
R2590	T12938	T12937	prep	to,common
R2591	T13088	T13089	punct	[,33
R2592	T13089	T13059	parataxis	33,were
R2593	T13090	T13089	punct	],33
R2594	T12939	T12940	preconj	both,calpain
R2595	T13091	T13051	punct	.,narrowed
R2596	T12940	T12938	pobj	calpain,to
R2597	T13093	T13094	prep	At,left
R2598	T13095	T13096	det	that,point
R2599	T12941	T12940	nmod	μ,calpain
R26	T696	T695	punct	", ",migration
R260	T950	T949	punct	-,calpain
R2600	T13096	T13093	pobj	point,At
R2601	T13097	T13094	punct	", ",left
R2602	T13098	T13094	nsubjpass	we,left
R2603	T12942	T12941	punct	-,μ
R2604	T13099	T13094	auxpass	were,left
R2605	T13100	T13094	prep	with,left
R2606	T13101	T13102	det	the,possibilities
R2607	T12943	T12941	cc	and,μ
R2608	T13102	T13100	pobj	possibilities,with
R2609	T13103	T13102	amod	last,possibilities
R261	T951	T947	acomp	vital,is
R2610	T13104	T13102	nummod	two,possibilities
R2611	T12944	T12941	conj	m,μ
R2612	T13105	T13106	mark	that,required
R2613	T13106	T13102	advcl	required,possibilities
R2614	T13107	T13108	det	either,calpain
R2615	T13108	T13106	nsubjpass	calpain,required
R2616	T13109	T13108	compound	m,calpain
R2617	T13110	T13108	punct	-,calpain
R2618	T13111	T13106	auxpass	was,required
R2619	T13112	T13106	advmod	specifically,required
R262	T952	T951	prep	for,vital
R2620	T13113	T13106	prep	during,required
R2621	T12945	T12940	punct	-,calpain
R2622	T13114	T13113	pobj	embryogenesis,during
R2623	T13115	T13106	punct	", ",required
R2624	T13116	T13106	cc	or,required
R2625	T13117	T13118	mark	that,was
R2626	T12946	T12923	punct	.,used
R2627	T13118	T13106	conj	was,required
R2628	T13119	T13120	det	either,isoform
R2629	T13120	T13118	nsubj	isoform,was
R263	T953	T952	pobj	development,for
R2630	T13121	T13120	advmod	alone,isoform
R2631	T13122	T13118	acomp	sufficient,was
R2632	T12948	T12949	nummod	Two,laboratories
R2633	T13123	T13122	prep	for,sufficient
R2634	T13124	T13123	pcomp	sustaining,for
R2635	T13125	T13126	amod	embryonic,viability
R2636	T12949	T12951	nsubj	laboratories,observed
R2637	T13126	T13124	dobj	viability,sustaining
R2638	T13127	T13094	punct	.,left
R2639	T13129	T13130	nsubj	We,report
R264	T954	T953	prep	of,development
R2640	T12950	T12949	amod	independent,laboratories
R2641	T13131	T13130	advmod	here,report
R2642	T13132	T13133	mark	that,die
R2643	T12952	T12953	amod	embryonic,lethality
R2644	T13133	T13130	ccomp	die,report
R2645	T13134	T13135	nmod	Capn2,mice
R2646	T13135	T13133	nsubj	mice,die
R2647	T13136	T13134	punct	-,Capn2
R2648	T13137	T13134	punct	/,Capn2
R2649	T13138	T13134	punct	-,Capn2
R265	T955	T956	det	the,embryo
R2650	T12953	T12951	dobj	lethality,observed
R2651	T13139	T13135	punct	", ",mice
R2652	T13140	T13141	dep	which,lack
R2653	T13141	T13135	relcl	lack,mice
R2654	T12954	T12951	prep	in,observed
R2655	T13142	T13143	det	the,subunit
R2656	T13143	T13141	dobj	subunit,lack
R2657	T13144	T13145	nmod	m,calpain
R2658	T12955	T12956	compound	Capn4,mice
R2659	T13145	T13143	nmod	calpain,subunit
R266	T956	T954	pobj	embryo,of
R2660	T13146	T13145	punct	-,calpain
R2661	T13147	T13143	amod	catalytic,subunit
R2662	T12956	T12954	pobj	mice,in
R2663	T13148	T13133	punct	", ",die
R2664	T13149	T13133	prep	at,die
R2665	T13150	T13151	det	the,stage
R2666	T12957	T12956	compound	knockout,mice
R2667	T13151	T13149	pobj	stage,at
R2668	T13152	T13151	compound	preimplantation,stage
R2669	T13153	T13151	prep	of,stage
R267	T957	T956	nmod	preimplantation,embryo
R2670	T13154	T13153	pobj	development,of
R2671	T13155	T13130	punct	.,report
R2672	T12958	T12951	punct	", ",observed
R2673	T13157	T13158	det	This,observation
R2674	T12959	T12960	dep	albeit,at
R2675	T12960	T12951	prep	at,observed
R2676	T13158	T13159	nsubj	observation,allows
R2677	T12961	T12962	amod	different,stages
R2678	T13160	T13161	nsubj	us,conclude
R2679	T13161	T13159	ccomp	conclude,allows
R268	T958	T956	amod	murine,embryo
R2680	T12962	T12960	pobj	stages,at
R2681	T13162	T13161	aux	to,conclude
R2682	T13163	T13161	advmod	now,conclude
R2683	T13164	T13165	mark	that,required
R2684	T12963	T12962	prep	of,stages
R2685	T13165	T13161	ccomp	required,conclude
R2686	T13166	T13167	compound	m,calpain
R2687	T12964	T12963	pobj	development,of
R2688	T13167	T13165	nsubjpass	calpain,required
R2689	T13168	T13167	punct	-,calpain
R269	T959	T932	punct	.,establishes
R2690	T13169	T13165	auxpass	is,required
R2691	T12965	T12966	punct	[,32
R2692	T13170	T13165	advmod	specifically,required
R2693	T13171	T13165	prep	during,required
R2694	T13172	T13171	pobj	embryogenesis,during
R2695	T12966	T12951	parataxis	32,observed
R2696	T13173	T13159	punct	.,allows
R2697	T12967	T12966	nummod	31,32
R2698	T13175	T13176	mark	Since,expected
R2699	T13176	T13185	advcl	expected,is
R27	T697	T698	compound	cell,cycle
R270	T676	T675	pobj	mice,in
R2700	T13177	T13178	amod	homozygous,disruption
R2701	T12968	T12966	punct	",",32
R2702	T13178	T13176	nsubjpass	disruption,expected
R2703	T12969	T12966	punct	],32
R2704	T13179	T13178	prep	of,disruption
R2705	T13180	T13181	det	the,gene
R2706	T13181	T13179	pobj	gene,of
R2707	T12970	T12951	punct	.,observed
R2708	T13182	T13181	compound	Capn4,gene
R2709	T13183	T13176	auxpass	was,expected
R2710	T12972	T12973	det	These,observations
R2711	T13184	T13176	advmod	also,expected
R2712	T13186	T13187	aux	to,abolish
R2713	T13187	T13176	xcomp	abolish,expected
R2714	T12973	T12974	nsubj	observations,supported
R2715	T13188	T13189	compound	m,calpain
R2716	T13189	T13191	compound	calpain,activity
R2717	T13190	T13189	punct	-,calpain
R2718	T12974	T12975	ccomp	supported,required
R2719	T13191	T13187	dobj	activity,abolish
R2720	T13192	T13185	punct	", ",is
R2721	T12976	T12977	det	the,hypothesis
R2722	T13193	T13194	det	this,result
R2723	T13194	T13185	nsubj	result,is
R2724	T12977	T12974	dobj	hypothesis,supported
R2725	T13195	T13185	prep	in,is
R2726	T12978	T12979	mark	that,required
R2727	T13196	T13195	pobj	agreement,in
R2728	T13197	T13196	prep	with,agreement
R2729	T13198	T13199	det	the,phenotype
R2730	T13199	T13197	pobj	phenotype,with
R2731	T13200	T13199	acl	presented,phenotype
R2732	T12979	T12977	acl	required,hypothesis
R2733	T13201	T13200	agent	by,presented
R2734	T13202	T13201	pobj	one,by
R2735	T13203	T13202	prep	of,one
R2736	T12980	T12981	det	the,subunit
R2737	T13204	T13205	det	the,lines
R2738	T13205	T13203	pobj	lines,of
R2739	T13299	T13300	det	the,cassette
R2740	T13206	T13205	nummod	two,lines
R2741	T13207	T13208	npadvmod	Capn4,targeted
R2742	T13208	T13205	amod	targeted,lines
R2743	T13209	T13210	prep	in,observed
R2744	T13300	T13298	pobj	cassette,of
R2745	T13210	T13205	relcl	observed,lines
R2746	T13211	T13209	pobj	which,in
R2747	T13212	T13213	compound	preimplantation,lethality
R2748	T13301	T13302	compound	PGK,Neo
R2749	T13213	T13210	nsubjpass	lethality,observed
R2750	T13214	T13213	prep	of,lethality
R2751	T13215	T13216	amod	null,embryos
R2752	T13216	T13214	pobj	embryos,of
R2753	T13302	T13300	compound	Neo,cassette
R2754	T13217	T13210	auxpass	was,observed
R2755	T13218	T13210	advmod	also,observed
R2756	T13219	T13220	punct	[,32
R2757	T13303	T13302	punct	-,Neo
R2758	T13304	T13297	prep	into,insertion
R2759	T13220	T13210	parataxis	32,observed
R2760	T13305	T13306	det	the,middle
R2761	T13221	T13220	punct	],32
R2762	T13222	T13185	punct	.,is
R2763	T13306	T13304	pobj	middle,into
R2764	T13224	T13225	det	The,survival
R2765	T13307	T13306	prep	of,middle
R2766	T13225	T13226	nsubj	survival,is
R2767	T13308	T13307	pobj	exon,of
R2768	T13227	T13225	prep	of,survival
R2769	T13228	T13229	nmod	Capn4,embryos
R2770	T13229	T13227	pobj	embryos,of
R2771	T13230	T13228	punct	-,Capn4
R2772	T13231	T13228	punct	/,Capn4
R2773	T13232	T13228	punct	-,Capn4
R2774	T13233	T13225	prep	from,survival
R2775	T13309	T13308	nummod	9,exon
R2776	T13234	T13235	det	the,line
R2777	T13235	T13233	pobj	line,from
R2778	T13236	T13235	amod	original,line
R2779	T13310	T13297	punct	", ",insertion
R2780	T13237	T13235	amod	targeted,line
R2781	T13238	T13235	acl	reported,line
R2782	T13311	T13312	dep	which,caused
R2783	T13239	T13238	agent	by,reported
R2784	T13240	T13239	pobj	Arthur,by
R2785	T13241	T13240	cc	and,Arthur
R2786	T13312	T13297	relcl	caused,insertion
R2787	T13242	T13240	conj	colleagues,Arthur
R2788	T13313	T13312	dobj	truncation,caused
R2789	T13314	T13313	prep	of,truncation
R2790	T13243	T13233	prep	to,from
R2791	T13244	T13243	pobj	mid-gestation,to
R2792	T13315	T13316	det	the,protein
R2793	T13245	T13246	advmod	more,difficult
R2794	T13246	T13226	acomp	difficult,is
R2795	T13316	T13314	pobj	protein,of
R2796	T13247	T13248	aux	to,reconcile
R2797	T13248	T13246	advcl	reconcile,difficult
R2798	T13249	T13250	punct	[,31
R2799	T13317	T13318	punct	[,31
R28	T698	T699	compound	cycle,progression
R2800	T13250	T13226	parataxis	31,is
R2801	T13251	T13250	punct	],31
R2802	T13318	T13312	parataxis	31,caused
R2803	T13252	T13226	punct	.,is
R2804	T13254	T13255	prep	In,seems
R2805	T13319	T13318	punct	],31
R2806	T13256	T13254	pobj	retrospect,In
R2807	T13320	T13280	punct	.,involved
R2808	T13257	T13255	punct	", ",seems
R2809	T13258	T13255	nsubj	it,seems
R2810	T13259	T13255	oprd	likely,seems
R2811	T13322	T13323	det	This,strategy
R2812	T13260	T13261	mark	that,represents
R2813	T13261	T13255	ccomp	represents,seems
R2814	T13262	T13263	det	the,line
R2815	T13263	T13261	nsubj	line,represents
R2816	T13323	T13324	nsubjpass	strategy,based
R2817	T13264	T13263	amod	latter,line
R2818	T13265	T13266	det	a,state
R2819	T13325	T13324	auxpass	was,based
R282	T3323	T3320	nummod	80,k
R2820	T13266	T13261	dobj	state,represents
R2821	T13267	T13266	amod	hypomorphic,state
R2822	T13268	T13266	punct	", ",state
R2823	T13269	T13270	advmod	rather,than
R2824	T13270	T13266	cc	than,state
R2825	T13271	T13272	det	a,null
R2826	T13272	T13266	conj	null,state
R2827	T13273	T13272	amod	true,null
R2828	T13274	T13255	punct	.,seems
R2829	T13326	T13324	prep	upon,based
R2830	T13276	T13277	det	The,strategy
R2831	T13277	T13280	nsubj	strategy,involved
R2832	T13327	T13328	amod	previous,studies
R2833	T13278	T13277	compound	Capn4,strategy
R2834	T13279	T13277	compound	targeting,strategy
R2835	T13328	T13326	pobj	studies,upon
R2836	T13281	T13277	acl	employed,strategy
R2837	T13282	T13281	agent	by,employed
R2838	T13283	T13282	pobj	Arthur,by
R2839	T13284	T13283	cc	and,Arthur
R2840	T13285	T13283	conj	colleagues,Arthur
R2841	T13329	T13330	compound	structure,function
R2842	T13286	T13280	xcomp	disrupting,involved
R2843	T13287	T13288	det	the,terminus
R2844	T13288	T13286	dobj	terminus,disrupting
R2845	T13330	T13328	compound	function,studies
R2846	T13289	T13288	compound	C,terminus
R2847	T13290	T13288	punct	-,terminus
R2848	T13291	T13288	prep	of,terminus
R2849	T13292	T13293	det	the,subunit
R2850	T13331	T13330	punct	/,function
R2851	T13293	T13291	pobj	subunit,of
R2852	T13294	T13293	nmod	calpain,subunit
R2853	T13332	T13328	acl	showing,studies
R2854	T13295	T13293	amod	small,subunit
R2855	T13296	T13286	agent	by,disrupting
R2856	T13297	T13296	pobj	insertion,by
R2857	T13333	T13334	mark	that,abolished
R2858	T13298	T13297	prep	of,insertion
R2859	T13334	T13332	ccomp	abolished,showing
R2860	T13335	T13334	nsubj	excision,abolished
R2861	T13336	T13335	prep	of,excision
R2862	T13405	T13406	det	the,time
R2863	T13406	T13404	pobj	time,in
R2864	T13337	T13338	det	the,acid
R2865	T13407	T13406	prep	of,time
R2866	T13408	T13407	pobj	lethality,of
R2867	T13409	T13406	prep	of,time
R2868	T13338	T13336	pobj	acid,of
R2869	T13410	T13411	det	these,lines
R287	T3324	T3320	punct	),k
R2870	T13339	T13340	npadvmod	C,terminal
R2871	T13340	T13338	amod	terminal,acid
R2872	T13411	T13409	pobj	lines,of
R2873	T13341	T13340	punct	-,terminal
R2874	T13412	T13411	nummod	two,lines
R2875	T13413	T13411	amod	targeted,lines
R2876	T13414	T13411	compound	Capn4,lines
R2877	T13415	T13401	aux	can,explained
R2878	T13416	T13401	auxpass	be,explained
R2879	T13417	T13401	agent	by,explained
R288	T3325	T3306	punct	", ",encoded
R2880	T13418	T13419	amod	different,extents
R2881	T13342	T13338	nummod	25,acid
R2882	T13419	T13417	pobj	extents,by
R2883	T13420	T13419	prep	of,extents
R2884	T13421	T13420	pobj	disruption,of
R2885	T13343	T13338	compound	amino,acid
R2886	T13422	T13397	punct	.,seems
R2887	T13344	T13338	nmod	residues,acid
R2888	T13424	T13425	det	The,allele
R2889	T13425	T13427	nsubj	allele,represents
R289	T3326	T3306	advmod	respectively,encoded
R2890	T13345	T13338	prep	of,acid
R2891	T13426	T13425	compound	Zimmerman,allele
R2892	T13346	T13347	det	the,subunit
R2893	T13428	T13427	advmod	probably,represents
R2894	T13429	T13430	det	a,genotype
R2895	T13430	T13427	dobj	genotype,represents
R2896	T13347	T13345	pobj	subunit,of
R2897	T13431	T13430	amod	true,genotype
R2898	T13432	T13430	amod	null,genotype
R2899	T13433	T13434	mark	while,is
R29	T699	T695	conj	progression,migration
R290	T3244	T3245	det	The,proteases
R2900	T13348	T13347	amod	small,subunit
R2901	T13434	T13427	advcl	is,represents
R2902	T13435	T13436	det	the,allele
R2903	T13436	T13434	nsubj	allele,is
R2904	T13349	T13350	compound	calpain,activity
R2905	T13437	T13436	compound	Arthur,allele
R2906	T13438	T13434	advmod	likely,is
R2907	T13439	T13440	det	a,mutation
R2908	T13350	T13334	dobj	activity,abolished
R2909	T13440	T13434	attr	mutation,is
R291	T3327	T3301	punct	", ",subunits
R2910	T13441	T13440	amod	hypomorphic,mutation
R2911	T13442	T13427	punct	.,represents
R2912	T13351	T13352	advmod	when,co-expressed
R2913	T13444	T13445	amod	Alternate,strategies
R2914	T13352	T13334	advcl	co-expressed,abolished
R2915	T13445	T13447	nsubjpass	strategies,shown
R2916	T13446	T13445	compound	targeting,strategies
R2917	T13353	T13352	prep	with,co-expressed
R2918	T13448	T13447	aux	have,shown
R2919	T13449	T13447	auxpass	been,shown
R292	T3245	T3254	nsubj	proteases,are
R2920	T13450	T13451	aux	to,yield
R2921	T13354	T13355	det	the,subunit
R2922	T13451	T13447	advcl	yield,shown
R2923	T13452	T13453	amod	different,phenotypes
R2924	T13453	T13451	dobj	phenotypes,yield
R2925	T13355	T13353	pobj	subunit,with
R2926	T13454	T13451	prep	in,yield
R2927	T13455	T13456	compound	gene,disruption
R2928	T13456	T13457	compound	disruption,studies
R2929	T13457	T13454	pobj	studies,in
R293	T3246	T3245	nummod	two,proteases
R2930	T13458	T13447	punct	.,shown
R2931	T13460	T13461	prep	For,used
R2932	T13356	T13357	nmod	m,calpain
R2933	T13462	T13460	pobj	example,For
R2934	T13463	T13461	punct	", ",used
R2935	T13464	T13465	nummod	three,strategies
R2936	T13465	T13461	nsubjpass	strategies,used
R2937	T13466	T13465	amod	different,strategies
R2938	T13357	T13355	nmod	calpain,subunit
R2939	T13467	T13465	compound	targeting,strategies
R294	T3328	T3329	dep	that,associate
R2940	T13468	T13461	auxpass	were,used
R2941	T13469	T13461	advmod	independently,used
R2942	T13358	T13357	punct	-,calpain
R2943	T13470	T13471	aux	to,disrupt
R2944	T13471	T13461	advcl	disrupt,used
R2945	T13359	T13355	amod	large,subunit
R2946	T13472	T13473	det	the,gene
R2947	T13473	T13471	dobj	gene,disrupt
R2948	T13474	T13475	amod	mixed,leukemia
R2949	T13475	T13473	nmod	leukemia,gene
R295	T3247	T3245	amod	ubiquitous,proteases
R2950	T13360	T13352	prep	in,co-expressed
R2951	T13476	T13475	nmod	lineage,leukemia
R2952	T13477	T13475	punct	(,leukemia
R2953	T13361	T13362	compound	E.,coli
R2954	T13478	T13475	appos	Mll,leukemia
R2955	T13479	T13473	punct	),gene
R2956	T13480	T13461	punct	.,used
R2957	T13362	T13360	pobj	coli,in
R2958	T13482	T13483	prep	In,perished
R2959	T13484	T13485	det	all,studies
R296	T3248	T3249	npadvmod	Ca2,dependent
R2960	T13363	T13364	punct	[,34
R2961	T13485	T13482	pobj	studies,In
R2962	T13486	T13485	nummod	three,studies
R2963	T13487	T13483	punct	", ",perished
R2964	T13364	T13334	parataxis	34,abolished
R2965	T13488	T13489	amod	homozygous,null
R2966	T13489	T13490	amod	null,embryos
R2967	T13490	T13483	nsubj	embryos,perished
R2968	T13365	T13364	punct	],34
R2969	T13491	T13483	prep	during,perished
R297	T3329	T3301	relcl	associate,subunits
R2970	T13492	T13491	pobj	embryogenesis,during
R2971	T13366	T13324	punct	.,based
R2972	T13493	T13491	punct	", ",during
R2973	T13494	T13491	cc	but,during
R2974	T13495	T13491	conj	at,during
R2975	T13496	T13497	amod	different,stages
R2976	T13368	T13369	prep	In,involved
R2977	T13497	T13495	pobj	stages,at
R2978	T13498	T13499	punct	(,E14.5
R2979	T13499	T13497	parataxis	E14.5,stages
R298	T3249	T3245	amod	dependent,proteases
R2980	T13500	T13499	nmod	E0.5,E14.5
R2981	T13501	T13499	punct	", ",E14.5
R2982	T13370	T13368	pobj	contrast,In
R2983	T13502	T13499	nmod	E10.5,E14.5
R2984	T13503	T13499	punct	", ",E14.5
R2985	T13504	T13499	punct	),E14.5
R2986	T13505	T13506	punct	[,35
R2987	T13506	T13483	parataxis	35,perished
R2988	T13371	T13369	punct	", ",involved
R2989	T13507	T13506	punct	],35
R299	T3250	T3248	punct	+,Ca2
R2990	T13508	T13483	punct	.,perished
R2991	T13372	T13373	det	the,strategy
R2992	T13510	T13511	det	The,variation
R2993	T13373	T13369	nsubj	strategy,involved
R2994	T13374	T13373	nmod	Capn4,strategy
R2995	T13375	T13373	amod	targeting,strategy
R2996	T13511	T13512	nsubjpass	variation,attributed
R2997	T13376	T13373	acl	employed,strategy
R2998	T13513	T13511	prep	in,variation
R2999	T13377	T13376	agent	by,employed
R3	T665	T666	compound	m,Calpain
R30	T700	T699	punct	", ",progression
R300	T3251	T3249	punct	-,dependent
R3000	T13514	T13513	pobj	phenotype,in
R3001	T13515	T13512	auxpass	was,attributed
R3002	T13516	T13512	prep	to,attributed
R3003	T13517	T13518	det	the,differences
R3004	T13378	T13377	pobj	Zimmerman,by
R3005	T13518	T13516	pobj	differences,to
R3006	T13519	T13518	prep	in,differences
R3007	T13520	T13519	pobj	degree,in
R3008	T13379	T13378	cc	and,Zimmerman
R3009	T13521	T13520	prep	of,degree
R301	T3330	T3329	prep	with,associate
R3010	T13522	T13521	pobj	function,of
R3011	T13523	T13520	prep	of,degree
R3012	T13380	T13378	conj	colleagues,Zimmerman
R3013	T13524	T13525	det	the,proteins
R3014	T13525	T13523	pobj	proteins,of
R3015	T13526	T13525	amod	truncated,proteins
R3016	T13381	T13382	det	a,deletion
R3017	T13527	T13525	acl	produced,proteins
R3018	T13528	T13527	prep	from,produced
R3019	T13529	T13530	det	the,alleles
R302	T3252	T3245	punct	", ",proteases
R3020	T13382	T13369	dobj	deletion,involved
R3021	T13530	T13528	pobj	alleles,from
R3022	T13531	T13530	compound	mutant,alleles
R3023	T13532	T13512	punct	.,attributed
R3024	T13383	T13384	advmod	much,more
R3025	T13534	T13535	det	A,effect
R3026	T13535	T13537	nsubj	effect,be
R3027	T13384	T13385	advmod	more,extensive
R3028	T13536	T13535	amod	similar,effect
R3029	T13538	T13537	aux	might,be
R303	T3253	T3245	compound	cysteine,proteases
R3030	T13385	T13382	amod	extensive,deletion
R3031	T13539	T13537	prep	at,be
R3032	T13540	T13539	pobj	work,at
R3033	T13541	T13537	prep	in,be
R3034	T13542	T13543	det	the,lines
R3035	T13543	T13541	pobj	lines,in
R3036	T13544	T13543	nummod	two,lines
R3037	T13386	T13382	prep	of,deletion
R3038	T13545	T13546	npadvmod	Capn4,transgenic
R3039	T13546	T13543	amod	transgenic,lines
R304	T3255	T3245	acl	known,proteases
R3040	T13547	T13537	punct	.,be
R3041	T13387	T13388	nmod	exons,4
R3042	T13549	T13550	nsubjpass	Efforts,made
R3043	T13388	T13386	pobj	4,of
R3044	T13551	T13550	auxpass	were,made
R3045	T13552	T13553	aux	to,detect
R3046	T13553	T13550	advcl	detect,made
R3047	T13389	T13388	prep	through,4
R3048	T13554	T13553	dobj	transcripts,detect
R3049	T13555	T13554	cc	or,transcripts
R305	T3331	T3332	det	a,subunit
R3050	T13390	T13389	pobj	8,through
R3051	T13556	T13557	compound	calpain,activities
R3052	T13557	T13554	conj	activities,transcripts
R3053	T13391	T13392	punct	[,32
R3054	T13558	T13554	acl	derived,transcripts
R3055	T13559	T13558	prep	from,derived
R3056	T13560	T13561	det	the,allele
R3057	T13561	T13559	pobj	allele,from
R3058	T13392	T13369	parataxis	32,involved
R3059	T13562	T13563	compound	Arthur,Capn4
R306	T3256	T3255	prep	as,known
R3060	T13563	T13561	compound	Capn4,allele
R3061	T13564	T13550	punct	.,made
R3062	T13393	T13392	punct	],32
R3063	T13566	T13567	det	This,allele
R3064	T13567	T13568	nsubj	allele,gave
R3065	T13394	T13369	punct	.,involved
R3066	T13569	T13568	dobj	rise,gave
R3067	T13396	T13397	nsubj	It,seems
R3068	T13570	T13568	prep	to,gave
R3069	T13571	T13572	amod	multiple,species
R307	T3257	T3258	compound	μ,calpain
R3070	T13572	T13570	pobj	species,to
R3071	T13573	T13572	compound	mRNA,species
R3072	T13398	T13397	advmod	now,seems
R3073	T13574	T13572	punct	", ",species
R3074	T13575	T13572	amod	detectable,species
R3075	T13576	T13575	prep	by,detectable
R3076	T13577	T13578	compound	RT,PCR
R3077	T13399	T13397	oprd	probable,seems
R3078	T13578	T13576	pobj	PCR,by
R3079	T13579	T13578	punct	-,PCR
R308	T3258	T3256	pobj	calpain,as
R3080	T13580	T13568	punct	", ",gave
R3081	T13400	T13401	mark	that,explained
R3082	T13581	T13568	advcl	reading,gave
R3083	T13582	T13581	advmod	through,reading
R3084	T13583	T13581	prep	from,reading
R3085	T13584	T13585	det	the,half
R3086	T13585	T13583	pobj	half,from
R3087	T13401	T13397	ccomp	explained,seems
R3088	T13586	T13585	amod	first,half
R3089	T13587	T13585	prep	of,half
R309	T3332	T3330	pobj	subunit,with
R3090	T13588	T13587	pobj	exon,of
R3091	T13402	T13403	det	the,difference
R3092	T13403	T13401	nsubjpass	difference,explained
R3093	T13589	T13588	nummod	9,exon
R3094	T13590	T13583	prep	to,from
R3095	T13404	T13403	prep	in,difference
R3096	T13591	T13592	advmod	at,two
R3097	T13592	T13594	nummod	two,sites
R3098	T13593	T13592	advmod	least,two
R3099	T13617	T13615	amod	small,subunits
R31	T701	T702	amod	degenerative,processes
R310	T3259	T3258	punct	-,calpain
R3100	T13594	T13590	pobj	sites,to
R3101	T13595	T13594	amod	different,sites
R3102	T13596	T13594	amod	cryptic,sites
R3103	T13597	T13594	compound	splice,sites
R3104	T13618	T13610	prep	with,give
R3105	T13598	T13594	prep	in,sites
R3106	T13599	T13600	det	the,sequence
R3107	T13619	T13620	quantmod	10,30
R3108	T13600	T13598	pobj	sequence,in
R3109	T13601	T13600	compound	PGK,sequence
R311	T3260	T3258	punct	(,calpain
R3110	T13602	T13600	compound	promoter,sequence
R3111	T13620	T13622	nummod	30,acids
R3112	T13603	T13604	punct	[,31
R3113	T13604	T13581	parataxis	31,reading
R3114	T13605	T13604	punct	],31
R3115	T13621	T13620	punct	–,30
R3116	T13606	T13568	punct	.,gave
R3117	T13608	T13609	det	These,transcripts
R3118	T13622	T13618	pobj	acids,with
R3119	T13609	T13610	nsubj	transcripts,give
R312	T3261	T3258	appos	calpain,calpain
R3120	T13611	T13610	aux	could,give
R3121	T13623	T13622	amod	inappropriate,acids
R3122	T13612	T13610	dobj	rise,give
R3123	T13613	T13610	prep	to,give
R3124	T13614	T13615	amod	defective,subunits
R3125	T13624	T13625	npadvmod	C,terminal
R3126	T13615	T13613	pobj	subunits,to
R3127	T13625	T13622	amod	terminal,acids
R3128	T13626	T13625	punct	-,terminal
R3129	T13616	T13615	nmod	calpain,subunits
R313	T3333	T3332	amod	common,subunit
R3130	T13627	T13622	punct	", ",acids
R3131	T13628	T13629	dep	which,be
R3132	T13629	T13622	relcl	be,acids
R3133	T13723	T13721	pobj	hands,in
R3134	T13630	T13629	aux	might,be
R3135	T13724	T13725	advmod	when,expressed
R3136	T13725	T13719	advcl	expressed,observed
R3137	T13726	T13727	nmod	calpain,subunits
R3138	T13727	T13725	nsubjpass	subunits,expressed
R3139	T13728	T13727	amod	large,subunits
R314	T3262	T3261	punct	-,calpain
R3140	T13729	T13725	auxpass	were,expressed
R3141	T13631	T13629	acomp	sufficient,be
R3142	T13730	T13725	advmod	alone,expressed
R3143	T13731	T13732	preconj	either,in
R3144	T13732	T13725	prep	in,expressed
R3145	T13632	T13633	aux	to,support
R3146	T13733	T13734	compound	E.,coli
R3147	T13734	T13732	pobj	coli,in
R3148	T13735	T13732	cc	or,in
R3149	T13633	T13631	xcomp	support,sufficient
R315	T3263	T3261	nummod	1,calpain
R3150	T13736	T13732	conj	in,in
R3151	T13737	T13738	amod	mammalian,cells
R3152	T13634	T13635	det	a,level
R3153	T13738	T13736	pobj	cells,in
R3154	T13739	T13740	punct	[,36
R3155	T13740	T13725	parataxis	36,expressed
R3156	T13635	T13633	dobj	level,support
R3157	T13741	T13740	punct	],36
R3158	T13742	T13655	punct	.,suggested
R3159	T13636	T13635	amod	low,level
R316	T3264	T3258	punct	),calpain
R3160	T13744	T13745	det	The,timing
R3161	T13745	T13747	nsubj	timing,be
R3162	T13637	T13635	prep	of,level
R3163	T13746	T13745	amod	different,timing
R3164	T13638	T13639	compound	calpain,activity
R3165	T13748	T13745	prep	of,timing
R3166	T13749	T13748	pobj	lethality,of
R3167	T13750	T13745	prep	in,timing
R3168	T13639	T13637	pobj	activity,of
R3169	T13751	T13752	det	the,knockouts
R317	T3334	T3335	nummod	28,kDa
R3170	T13752	T13750	pobj	knockouts,in
R3171	T13753	T13752	compound	Capn4,knockouts
R3172	T13640	T13610	punct	.,give
R3173	T13754	T13747	aux	might,be
R3174	T13755	T13747	advmod	also,be
R3175	T13756	T13757	det	a,consequence
R3176	T13642	T13643	advmod	However,give
R3177	T13643	T13655	ccomp	give,suggested
R3178	T13757	T13747	attr	consequence,be
R3179	T13758	T13757	prep	of,consequence
R318	T3265	T3258	cc	and,calpain
R3180	T13644	T13643	punct	", ",give
R3181	T13759	T13760	det	the,backgrounds
R3182	T13760	T13758	pobj	backgrounds,of
R3183	T13761	T13760	amod	different,backgrounds
R3184	T13645	T13643	nsubj	expression,give
R3185	T13762	T13760	amod	genetic,backgrounds
R3186	T13763	T13760	prep	of,backgrounds
R3187	T13764	T13765	det	the,lines
R3188	T13765	T13763	pobj	lines,of
R3189	T13766	T13765	nummod	two,lines
R319	T3335	T3332	nmod	kDa,subunit
R3190	T13646	T13645	prep	of,expression
R3191	T13767	T13765	amod	transgenic,lines
R3192	T13768	T13757	punct	", ",consequence
R3193	T13769	T13770	dep	which,observed
R3194	T13647	T13646	pobj	calpains,of
R3195	T13770	T13757	relcl	observed,consequence
R3196	T13771	T13770	aux	has,observed
R3197	T13772	T13770	auxpass	been,observed
R3198	T13648	T13647	prep	with,calpains
R3199	T13773	T13774	aux	to,influence
R32	T702	T699	conj	processes,progression
R320	T3266	T3267	compound	m,calpain
R3200	T13774	T13770	advcl	influence,observed
R3201	T13649	T13650	det	these,subunits
R3202	T13775	T13776	det	the,phenotype
R3203	T13776	T13774	dobj	phenotype,influence
R3204	T13777	T13776	prep	of,phenotype
R3205	T13650	T13648	pobj	subunits,with
R3206	T13778	T13779	amod	transgenic,mice
R3207	T13779	T13777	pobj	mice,of
R3208	T13651	T13650	amod	modified,subunits
R3209	T13780	T13774	prep	on,influence
R321	T3267	T3258	conj	calpain,calpain
R3210	T13781	T13782	det	a,number
R3211	T13782	T13780	pobj	number,on
R3212	T13652	T13650	amod	small,subunits
R3213	T13783	T13782	prep	of,number
R3214	T13784	T13783	pobj	occasions,of
R3215	T13785	T13786	punct	[,37
R3216	T13653	T13643	aux	did,give
R3217	T13786	T13774	parataxis	37,influence
R3218	T13787	T13786	punct	],37
R3219	T13654	T13643	neg	not,give
R322	T3268	T3267	punct	-,calpain
R3220	T13788	T13747	punct	.,be
R3221	T13790	T13791	nsubj	One,been
R3222	T13656	T13643	dobj	rise,give
R3223	T13792	T13790	prep	of,One
R3224	T13793	T13794	det	the,questions
R3225	T13794	T13792	pobj	questions,of
R3226	T13657	T13643	prep	to,give
R3227	T13795	T13794	amod	enduring,questions
R3228	T13796	T13794	prep	in,questions
R3229	T13797	T13798	compound	calpain,research
R323	T3269	T3267	punct	(,calpain
R3230	T13658	T13659	det	any,activity
R3231	T13798	T13796	pobj	research,in
R3232	T13799	T13791	aux	has,been
R3233	T13800	T13801	mark	whether,possess
R3234	T13659	T13657	pobj	activity,to
R3235	T13801	T13791	ccomp	possess,been
R3236	T13802	T13803	det	the,isoforms
R3237	T13803	T13801	nsubj	isoforms,possess
R3238	T13660	T13659	amod	detectable,activity
R3239	T13804	T13803	nummod	two,isoforms
R324	T3270	T3267	appos	capain,calpain
R3240	T13805	T13803	amod	ubiquitous,isoforms
R3241	T13806	T13803	punct	", ",isoforms
R3242	T13807	T13808	nmod	μ,calpain
R3243	T13808	T13803	appos	calpain,isoforms
R3244	T13661	T13659	compound	calpain,activity
R3245	T13809	T13807	punct	-,μ
R3246	T13810	T13807	cc	and,μ
R3247	T13811	T13807	conj	m,μ
R3248	T13812	T13808	punct	-,calpain
R3249	T13662	T13663	advmod	when,expressed
R325	T3271	T3270	punct	-,capain
R3250	T13813	T13801	punct	", ",possess
R3251	T13814	T13815	amod	distinct,roles
R3252	T13663	T13643	advcl	expressed,give
R3253	T13815	T13801	dobj	roles,possess
R3254	T13816	T13817	advmod	in,vivo
R3255	T13817	T13815	amod	vivo,roles
R3256	T13664	T13663	prep	in,expressed
R3257	T13818	T13791	punct	.,been
R3258	T13820	T13821	det	The,enzymes
R3259	T13665	T13666	compound	E.,coli
R326	T3272	T3270	nummod	2,capain
R3260	T13821	T13823	nsubj	enzymes,share
R3261	T13822	T13821	nummod	two,enzymes
R3262	T13666	T13664	pobj	coli,in
R3263	T13824	T13825	nummod	62,%
R3264	T13825	T13826	compound	%,identity
R3265	T13667	T13643	punct	", ",give
R3266	T13826	T13823	dobj	identity,share
R3267	T13827	T13826	compound	sequence,identity
R3268	T13828	T13823	cc	and,share
R3269	T13668	T13669	mark	although,has
R327	T3273	T3254	punct	),are
R3270	T13669	T13643	advcl	has,give
R3271	T13670	T13671	poss	their,functionality
R3272	T13829	T13823	conj	are,share
R3273	T13830	T13831	advmod	very,similar
R3274	T13671	T13669	nsubj	functionality,has
R3275	T13831	T13829	acomp	similar,are
R3276	T13832	T13829	prep	in,are
R3277	T13833	T13834	poss	their,structure
R3278	T13672	T13671	prep	in,functionality
R3279	T13834	T13832	pobj	structure,in
R328	T3274	T3254	punct	", ",are
R3280	T13835	T13834	cc	and,structure
R3281	T13673	T13674	amod	mammalian,cells
R3282	T13836	T13834	conj	biochemistry,structure
R3283	T13837	T13823	punct	.,share
R3284	T13674	T13672	pobj	cells,in
R3285	T13839	T13840	advmod	Notably,cleave
R3286	T13675	T13669	advmod	yet,has
R3287	T13841	T13840	punct	", ",cleave
R3288	T13842	T13840	nsubj	they,cleave
R3289	T13843	T13840	advmod	essentially,cleave
R329	T3275	T3276	det	the,members
R3290	T13676	T13677	aux	to,determined
R3291	T13677	T13669	xcomp	determined,has
R3292	T13844	T13845	det	the,set
R3293	T13845	T13840	dobj	set,cleave
R3294	T13678	T13677	auxpass	be,determined
R3295	T13846	T13845	amod	same,set
R3296	T13847	T13845	prep	of,set
R3297	T13848	T13847	pobj	substrates,of
R3298	T13679	T13680	punct	(,J.S.
R3299	T13849	T13850	advmod	in,vitro
R33	T703	T702	cc	and,processes
R330	T3276	T3254	attr	members,are
R3300	T13850	T13840	advmod	vitro,cleave
R3301	T13680	T13669	meta	J.S.,has
R3302	T13851	T13840	punct	", ",cleave
R3303	T13852	T13840	advcl	suggesting,cleave
R3304	T13681	T13680	nmod	Elce,J.S.
R3305	T13853	T13854	mark	that,have
R3306	T13854	T13852	ccomp	have,suggesting
R3307	T13855	T13854	nsubj	they,have
R3308	T13682	T13680	punct	", ",J.S.
R3309	T13856	T13857	det	the,potential
R331	T3277	T3276	compound	founding,members
R3310	T13857	T13854	dobj	potential,have
R3311	T13683	T13680	amod	unpublished,J.S.
R3312	T13858	T13859	aux	to,carry
R3313	T13859	T13857	acl	carry,potential
R3314	T13860	T13859	prt	out,carry
R3315	T13684	T13680	nmod	work,J.S.
R3316	T13861	T13862	det	the,functions
R3317	T13862	T13859	dobj	functions,carry
R3318	T13863	T13862	amod	same,functions
R3319	T13685	T13680	punct	),J.S.
R332	T3278	T3276	prep	of,members
R3320	T13864	T13865	advmod	in,vivo
R3321	T13865	T13859	advmod	vivo,carry
R3322	T13686	T13655	nsubjpass	It,suggested
R3323	T13866	T13840	punct	.,cleave
R3324	T13687	T13655	aux	has,suggested
R3325	T13868	T13869	prep	On,is
R3326	T13688	T13655	advmod	also,suggested
R3327	T13870	T13871	det	the,hand
R3328	T13871	T13868	pobj	hand,On
R3329	T13872	T13871	amod	other,hand
R333	T3336	T3332	amod	regulatory,subunit
R3330	T13689	T13655	auxpass	been,suggested
R3331	T13873	T13869	punct	", ",is
R3332	T13874	T13875	mark	since,require
R3333	T13875	T13869	advcl	require,is
R3334	T13690	T13691	mark	that,provide
R3335	T13876	T13875	nsubj	they,require
R3336	T13877	T13878	amod	different,amounts
R3337	T13878	T13875	dobj	amounts,require
R3338	T13691	T13655	ccomp	provide,suggested
R3339	T13879	T13878	prep	of,amounts
R334	T3279	T3280	det	a,family
R3340	T13880	T13879	pobj	Ca2,of
R3341	T13881	T13880	punct	+,Ca2
R3342	T13692	T13693	nmod	calpain,subunits
R3343	T13882	T13875	prep	for,require
R3344	T13883	T13884	nmod	in,vitro
R3345	T13884	T13885	nmod	vitro,activation
R3346	T13885	T13882	pobj	activation,for
R3347	T13886	T13869	punct	", ",is
R3348	T13693	T13691	nsubj	subunits,provide
R3349	T13887	T13869	nsubj	it,is
R335	T3280	T3278	pobj	family,of
R3350	T13888	T13869	acomp	possible,is
R3351	T13889	T13890	mark	that,regulated
R3352	T13694	T13693	amod	large,subunits
R3353	T13890	T13869	ccomp	regulated,is
R3354	T13891	T13892	det	the,isoforms
R3355	T13695	T13693	advmod	alone,subunits
R3356	T13892	T13890	nsubjpass	isoforms,regulated
R3357	T13893	T13892	nummod	two,isoforms
R3358	T13696	T13691	aux	might,provide
R3359	T13894	T13890	auxpass	are,regulated
R336	T3281	T3280	compound	gene,family
R3360	T13895	T13890	advmod	differentially,regulated
R3361	T13896	T13890	prep	inside,regulated
R3362	T13697	T13698	det	some,activity
R3363	T13897	T13896	pobj	cells,inside
R3364	T13898	T13869	punct	.,is
R3365	T13698	T13691	dobj	activity,provide
R3366	T13900	T13901	nsubj	It,is
R3367	T13699	T13691	prep	in,provide
R3368	T13902	T13901	advmod	now,is
R3369	T13903	T13901	acomp	clear,is
R337	T3282	T3280	acl	comprising,family
R3370	T13904	T13901	punct	", ",is
R3371	T13700	T13701	amod	eukaryotic,cells
R3372	T13905	T13901	prep	from,is
R3373	T13906	T13907	det	the,work
R3374	T13907	T13905	pobj	work,from
R3375	T13701	T13699	pobj	cells,in
R3376	T13908	T13909	compound	gene,targeting
R3377	T13909	T13907	compound	targeting,work
R3378	T13910	T13907	acl	done,work
R3379	T13702	T13691	punct	", ",provide
R338	T3337	T3332	acl	encoded,subunit
R3380	T13911	T13910	prep	in,done
R3381	T13912	T13911	pobj	mice,in
R3382	T13703	T13704	mark	although,appears
R3383	T13913	T13901	punct	", ",is
R3384	T13914	T13915	mark	that,have
R3385	T13915	T13901	ccomp	have,is
R3386	T13704	T13691	advcl	appears,provide
R3387	T13916	T13917	nmod	μ,calpain
R3388	T13917	T13915	nsubj	calpain,have
R3389	T13918	T13916	punct	-,μ
R339	T3283	T3284	nummod	13,genes
R3390	T13705	T13706	det	the,knockout
R3391	T13919	T13916	cc	and,μ
R3392	T13920	T13916	conj	m,μ
R3393	T13921	T13917	punct	-,calpain
R3394	T13706	T13704	nsubj	knockout,appears
R3395	T13922	T13923	det	some,roles
R3396	T13923	T13915	dobj	roles,have
R3397	T13924	T13923	amod	distinct,roles
R3398	T13925	T13923	amod	physiological,roles
R3399	T13926	T13915	punct	", ",have
R34	T704	T705	compound	cell,death
R340	T3284	T3282	dobj	genes,comprising
R3400	T13707	T13706	nmod	Zimmerman,knockout
R3401	T13927	T13928	advmod	at,least
R3402	T13928	T13929	advmod	least,during
R3403	T13708	T13709	advmod	et,al.
R3404	T13929	T13915	prep	during,have
R3405	T13709	T13707	advmod	al.,Zimmerman
R3406	T13930	T13929	pobj	embryogenesis,during
R3407	T13931	T13901	punct	.,is
R3408	T13710	T13711	aux	to,exclude
R3409	T13933	T13934	mark	As,noted
R341	T3285	T3282	prep	in,comprising
R3410	T13711	T13704	xcomp	exclude,appears
R3411	T13934	T13935	advcl	noted,affect
R3412	T13712	T13713	det	that,possibility
R3413	T13713	T13711	dobj	possibility,exclude
R3414	T13936	T13935	punct	", ",affect
R3415	T13714	T13691	punct	", ",provide
R3416	T13937	T13938	mark	whereas,die
R3417	T13938	T13935	advcl	die,affect
R3418	T13715	T13691	cc	and,provide
R3419	T13939	T13940	npadvmod	Capn2,null
R342	T3338	T3337	agent	by,encoded
R3420	T13940	T13941	amod	null,embryos
R3421	T13941	T13938	nsubj	embryos,die
R3422	T13716	T13717	det	no,activity
R3423	T13942	T13941	amod	murine,embryos
R3424	T13943	T13944	amod	prior,to
R3425	T13717	T13719	nsubjpass	activity,observed
R3426	T13944	T13938	prep	to,die
R3427	T13945	T13944	pobj	implantation,to
R3428	T13946	T13935	punct	", ",affect
R3429	T13718	T13717	compound	calpain,activity
R343	T3286	T3285	pobj	mammals,in
R3430	T13947	T13948	amod	homozygous,disruption
R3431	T13948	T13935	nsubj	disruption,affect
R3432	T13949	T13948	prep	of,disruption
R3433	T13719	T13691	conj	observed,provide
R3434	T13950	T13951	det	the,gene
R3435	T13951	T13949	pobj	gene,of
R3436	T13720	T13719	auxpass	was,observed
R3437	T13952	T13953	nmod	μ,calpain
R3438	T13953	T13951	nmod	calpain,gene
R3439	T13954	T13953	punct	-,calpain
R344	T3287	T3288	punct	[,1
R3440	T13721	T13719	prep	in,observed
R3441	T13955	T13951	amod	large,gene
R3442	T13956	T13951	compound	subunit,gene
R3443	T13722	T13723	poss	our,hands
R3444	T13957	T13951	punct	", ",gene
R3445	T13958	T13951	appos	Capn1,gene
R3446	T13959	T13935	punct	", ",affect
R3447	T13960	T13935	aux	did,affect
R3448	T13961	T13935	neg	not,affect
R3449	T13962	T13963	det	the,viability
R345	T3288	T3254	parataxis	1,are
R3450	T14041	T14040	amod	relative,amounts
R3451	T13963	T13935	dobj	viability,affect
R3452	T13964	T13963	prep	of,viability
R3453	T13965	T13964	pobj	mice,of
R3454	T13966	T13967	punct	[,33
R3455	T13967	T13935	parataxis	33,affect
R3456	T13968	T13967	punct	],33
R3457	T14042	T14013	punct	.,are
R3458	T13969	T13935	punct	.,affect
R3459	T13971	T13972	det	The,phenotype
R346	T3339	T3340	det	the,gene
R3460	T14044	T14045	nsubj	Platelets,contain
R3461	T13972	T13974	nsubj	phenotype,involved
R3462	T13973	T13972	amod	principal,phenotype
R3463	T14046	T14044	cc	and,Platelets
R3464	T13975	T13972	acl	observed,phenotype
R3465	T13976	T13975	prep	as,observed
R3466	T13977	T13978	det	a,result
R3467	T13978	T13976	pobj	result,as
R3468	T14047	T14044	conj	erythrocytes,Platelets
R3469	T13979	T13978	prep	of,result
R347	T3289	T3290	punct	-,3
R3470	T13980	T13981	compound	Capn1,deficiency
R3471	T13981	T13979	pobj	deficiency,of
R3472	T14048	T14049	amod	abundant,activity
R3473	T13982	T13983	det	a,disturbance
R3474	T13983	T13974	dobj	disturbance,involved
R3475	T13984	T13983	prep	in,disturbance
R3476	T14049	T14045	dobj	activity,contain
R3477	T13985	T13986	compound	platelet,function
R3478	T13986	T13984	pobj	function,in
R3479	T13987	T13983	punct	", ",disturbance
R348	T3290	T3288	prep	3,1
R3480	T14050	T14051	compound	μ,calpain
R3481	T13988	T13989	advmod	possibly,at
R3482	T13989	T13983	prep	at,disturbance
R3483	T13990	T13991	det	the,level
R3484	T14051	T14049	compound	calpain,activity
R3485	T13991	T13989	pobj	level,at
R3486	T13992	T13991	prep	of,level
R3487	T13993	T13994	det	the,phosphorylation
R3488	T14052	T14051	punct	-,calpain
R3489	T13994	T13992	pobj	phosphorylation,of
R349	T3291	T3288	punct	],1
R3490	T13995	T13994	compound	tyrosine,phosphorylation
R3491	T13996	T13994	prep	of,phosphorylation
R3492	T14053	T14045	punct	", ",contain
R3493	T13997	T13998	amod	certain,proteins
R3494	T13998	T13996	pobj	proteins,of
R3495	T13999	T13998	acl	involved,proteins
R3496	T14054	T14055	mark	while,is
R3497	T14055	T14045	advcl	is,contain
R3498	T14000	T13999	prep	in,involved
R3499	T14001	T14002	compound	platelet,activation
R35	T705	T702	conj	death,processes
R350	T3340	T3338	pobj	gene,by
R3500	T14002	T14000	pobj	activation,in
R3501	T14003	T13974	punct	.,involved
R3502	T14056	T14057	compound	m,calpain
R3503	T14005	T14006	preconj	Both,activities
R3504	T14006	T14013	nsubj	activities,are
R3505	T14057	T14059	compound	calpain,activity
R3506	T14007	T14008	nmod	μ,calpain
R3507	T14008	T14006	compound	calpain,activities
R3508	T14009	T14007	punct	-,μ
R3509	T14058	T14057	punct	-,calpain
R351	T3292	T3254	punct	.,are
R3510	T14010	T14007	cc	and,μ
R3511	T14011	T14007	conj	m,μ
R3512	T14012	T14008	punct	-,calpain
R3513	T14059	T14055	nsubj	activity,is
R3514	T14014	T14013	acomp	present,are
R3515	T14060	T14061	advmod	barely,detectable
R3516	T14015	T14013	prep	in,are
R3517	T14061	T14055	acomp	detectable,is
R3518	T14062	T14045	punct	.,contain
R3519	T14016	T14017	amod	most,cells
R352	T3341	T3340	compound	Capn4,gene
R3520	T14017	T14015	pobj	cells,in
R3521	T14018	T14017	amod	mammalian,cells
R3522	T14064	T14065	nsubj	Compensation,is
R3523	T14019	T14013	punct	", ",are
R3524	T14020	T14021	mark	although,provide
R3525	T14066	T14064	prep	for,Compensation
R3526	T14021	T14013	advcl	provide,are
R3527	T14022	T14023	det	the,data
R3528	T14023	T14021	nsubj	data,provide
R3529	T14024	T14023	amod	published,data
R353	T3294	T3295	nsubj	Both,are
R3530	T14067	T14068	compound	μ,calpain
R3531	T14025	T14023	punct	", ",data
R3532	T14026	T14023	prep	owing,data
R3533	T14027	T14026	prep	to,owing
R3534	T14068	T14070	compound	calpain,deficiency
R3535	T14028	T14027	pobj	weaknesses,to
R3536	T14029	T14028	prep	in,weaknesses
R3537	T14030	T14031	det	the,methodology
R3538	T14069	T14068	punct	-,calpain
R3539	T14031	T14029	pobj	methodology,in
R354	T3296	T3297	amod	heterodimeric,enzymes
R3540	T14032	T14031	amod	available,methodology
R3541	T14033	T14021	punct	", ",provide
R3542	T14070	T14066	pobj	deficiency,for
R3543	T14034	T14021	aux	do,provide
R3544	T14035	T14021	neg	not,provide
R3545	T14036	T14037	amod	reliable,estimates
R3546	T14071	T14064	prep	by,Compensation
R3547	T14037	T14021	dobj	estimates,provide
R3548	T14038	T14037	prep	of,estimates
R3549	T14039	T14040	poss	their,amounts
R355	T3342	T3295	punct	.,are
R3550	T14072	T14073	compound	m,calpain
R3551	T14040	T14038	pobj	amounts,of
R3552	T14073	T14071	pobj	calpain,by
R3553	T14074	T14073	punct	-,calpain
R3554	T14075	T14065	advmod	therefore,is
R3555	T14147	T14145	punct	-,Capn2
R3556	T14076	T14077	advmod	less,likely
R3557	T14148	T14145	punct	/,Capn2
R3558	T14149	T14145	punct	-,Capn2
R3559	T14077	T14065	acomp	likely,is
R356	T3297	T3295	attr	enzymes,are
R3560	T14150	T14151	mark	that,is
R3561	T14151	T14140	ccomp	is,demonstrates
R3562	T14078	T14077	prep	in,likely
R3563	T14152	T14153	compound	m,calpain
R3564	T14153	T14155	compound	calpain,activity
R3565	T14154	T14153	punct	-,calpain
R3566	T14079	T14078	pobj	platelets,in
R3567	T14155	T14151	nsubj	activity,is
R3568	T14156	T14151	acomp	essential,is
R3569	T14157	T14156	prep	for,essential
R357	T3298	T3297	acl	consisting,enzymes
R3570	T14080	T14079	cc	and,platelets
R3571	T14158	T14159	amod	embryonic,development
R3572	T14159	T14157	pobj	development,for
R3573	T14160	T14159	prep	beyond,development
R3574	T14081	T14079	conj	erythrocytes,platelets
R3575	T14161	T14162	det	the,stage
R3576	T14162	T14160	pobj	stage,beyond
R3577	T14082	T14077	prep	than,likely
R3578	T14163	T14164	nummod	8,cell
R3579	T14164	T14162	compound	cell,stage
R358	T3299	T3298	prep	of,consisting
R3580	T14165	T14164	punct	-,cell
R3581	T14083	T14082	prep	in,than
R3582	T14166	T14140	punct	.,demonstrates
R3583	T14168	T14169	nsubj	It,follows
R3584	T14084	T14085	amod	other,types
R3585	T14170	T14171	mark	that,are
R3586	T14171	T14169	ccomp	are,follows
R3587	T14085	T14083	pobj	types,in
R3588	T14172	T14173	advmod	at,least
R3589	T14173	T14174	advmod	least,functions
R359	T3344	T3345	det	The,subunits
R3590	T14086	T14085	compound	cell,types
R3591	T14174	T14171	nsubj	functions,are
R3592	T14175	T14174	det	some,functions
R3593	T14176	T14174	prep	of,functions
R3594	T14087	T14065	punct	.,is
R3595	T14177	T14178	compound	μ,calpain
R3596	T14178	T14176	pobj	calpain,of
R3597	T14179	T14178	punct	-,calpain
R3598	T14089	T14090	prep	As,is
R3599	T14180	T14178	cc	and,calpain
R36	T706	T684	punct	.,are
R360	T3300	T3301	amod	distinct,subunits
R3600	T14181	T14182	compound	m,calpain
R3601	T14182	T14178	conj	calpain,calpain
R3602	T14183	T14182	punct	-,calpain
R3603	T14184	T14171	acomp	distinct,are
R3604	T14185	T14169	punct	.,follows
R3605	T14091	T14092	det	a,result
R3606	T14187	T14188	det	The,cause
R3607	T14092	T14089	pobj	result,As
R3608	T14188	T14190	nsubjpass	cause,clarified
R3609	T14189	T14188	amod	underlying,cause
R361	T3301	T3299	pobj	subunits,of
R3610	T14093	T14090	punct	", ",is
R3611	T14191	T14188	prep	of,cause
R3612	T14192	T14193	det	the,lethality
R3613	T14193	T14191	pobj	lethality,of
R3614	T14094	T14090	nsubj	it,is
R3615	T14194	T14193	compound	preimplantation,lethality
R3616	T14195	T14188	prep	in,cause
R3617	T14196	T14197	nmod	Capn2,embryos
R3618	T14095	T14090	neg	not,is
R3619	T14197	T14195	pobj	embryos,in
R362	T3302	T3303	nummod	80,kDa
R3620	T14198	T14196	punct	-,Capn2
R3621	T14096	T14090	acomp	possible,is
R3622	T14199	T14196	punct	/,Capn2
R3623	T14200	T14196	punct	-,Capn2
R3624	T14201	T14190	aux	has,clarified
R3625	T14097	T14098	aux	to,determine
R3626	T14202	T14190	neg	not,clarified
R3627	T14203	T14190	advmod	yet,clarified
R3628	T14204	T14190	auxpass	been,clarified
R3629	T14098	T14090	xcomp	determine,is
R363	T3303	T3301	nmod	kDa,subunits
R3630	T14205	T14190	punct	.,clarified
R3631	T14099	T14100	mark	whether,is
R3632	T14207	T14208	det	The,embryos
R3633	T14208	T14212	nsubj	embryos,present
R3634	T14209	T14208	nummod	two,embryos
R3635	T14100	T14098	advcl	is,determine
R3636	T14210	T14211	amod	homozygous,null
R3637	T14211	T14208	amod	null,embryos
R3638	T14101	T14102	det	the,absence
R3639	T14213	T14208	acl	identified,embryos
R364	T3345	T3355	nsubj	subunits,share
R3640	T14214	T14213	prep	at,identified
R3641	T14215	T14216	det	the,stage
R3642	T14102	T14100	nsubj	absence,is
R3643	T14216	T14214	pobj	stage,at
R3644	T14217	T14218	nummod	8,cell
R3645	T14218	T14216	compound	cell,stage
R3646	T14103	T14102	prep	of,absence
R3647	T14219	T14218	punct	-,cell
R3648	T14220	T14212	aux	did,present
R3649	T14221	T14212	neg	not,present
R365	T3304	T3301	amod	catalytic,subunits
R3650	T14104	T14105	amod	marked,phenotype
R3651	T14222	T14223	det	any,defects
R3652	T14223	T14212	dobj	defects,present
R3653	T14105	T14103	pobj	phenotype,of
R3654	T14224	T14223	amod	obvious,defects
R3655	T14225	T14223	amod	morphological,defects
R3656	T14226	T14212	punct	.,present
R3657	T14228	T14229	advmod	However,is
R3658	T14106	T14102	prep	in,absence
R3659	T14230	T14229	punct	", ",is
R366	T3305	T3301	punct	", ",subunits
R3660	T14231	T14232	det	the,fact
R3661	T14232	T14229	nsubj	fact,is
R3662	T14107	T14108	nmod	Capn1,animals
R3663	T14233	T14234	mark	that,proved
R3664	T14234	T14232	acl	proved,fact
R3665	T14235	T14236	advmod	only,embryos
R3666	T14108	T14106	pobj	animals,in
R3667	T14236	T14234	nsubj	embryos,proved
R3668	T14237	T14238	quantmod	two,90
R3669	T14238	T14236	nummod	90,embryos
R367	T3346	T3347	nmod	μ,k
R3670	T14109	T14107	punct	-,Capn1
R3671	T14239	T14238	quantmod	out,90
R3672	T14240	T14238	quantmod	of,90
R3673	T14241	T14242	advmod	successfully,genotyped
R3674	T14110	T14107	punct	/,Capn1
R3675	T14242	T14236	amod	genotyped,embryos
R3676	T14243	T14236	amod	pre-implantation,embryos
R3677	T14244	T14245	aux	to,be
R3678	T14111	T14107	punct	-,Capn1
R3679	T14245	T14234	xcomp	be,proved
R368	T3347	T3345	nmod	k,subunits
R3680	T14246	T14245	attr	Capn2,be
R3681	T14247	T14246	punct	-,Capn2
R3682	T14112	T14100	prep	due,is
R3683	T14248	T14246	punct	/,Capn2
R3684	T14249	T14246	punct	-,Capn2
R3685	T14113	T14112	pcomp	to,due
R3686	T14250	T14229	prep	in,is
R3687	T14251	T14250	pobj	itself,in
R3688	T14252	T14229	acomp	revealing,is
R3689	T14114	T14115	det	a,affect
R369	T3306	T3301	acl	encoded,subunits
R3690	T14115	T14112	pobj	affect,due
R3691	T14116	T14115	amod	compensatory,affect
R3692	T14253	T14229	punct	.,is
R3693	T14117	T14115	prep	by,affect
R3694	T14255	T14256	compound	Preimplantation,lethality
R3695	T14256	T14257	nsubjpass	lethality,attributed
R3696	T14118	T14119	det	the,activity
R3697	T14258	T14256	acl	resulting,lethality
R3698	T14259	T14258	prep	from,resulting
R3699	T14119	T14117	pobj	activity,by
R37	T708	T709	det	These,enzymes
R370	T3307	T3306	agent	by,encoded
R3700	T14260	T14261	compound	gene,knockouts
R3701	T14261	T14259	pobj	knockouts,from
R3702	T14120	T14119	amod	remaining,activity
R3703	T14262	T14257	aux	can,attributed
R3704	T14263	T14257	advmod	typically,attributed
R3705	T14264	T14257	auxpass	be,attributed
R3706	T14265	T14257	prep	to,attributed
R3707	T14266	T14267	nummod	two,causes
R3708	T14267	T14265	pobj	causes,to
R3709	T14121	T14122	compound	m,calpain
R371	T3348	T3346	punct	-,μ
R3710	T14268	T14267	amod	general,causes
R3711	T14269	T14257	punct	.,attributed
R3712	T14122	T14119	compound	calpain,activity
R3713	T14271	T14272	prep	In,incurred
R3714	T14273	T14274	det	some,cases
R3715	T14123	T14122	punct	-,calpain
R3716	T14274	T14271	pobj	cases,In
R3717	T14275	T14272	punct	", ",incurred
R3718	T14124	T14100	punct	", ",is
R3719	T14276	T14272	nsubjpass	defects,incurred
R372	T3308	T3309	det	the,genes
R3720	T14277	T14272	auxpass	are,incurred
R3721	T14278	T14272	prep	in,incurred
R3722	T14125	T14100	cc	or,is
R3723	T14126	T14127	mark	whether,are
R3724	T14279	T14280	det	the,program
R3725	T14127	T14100	conj	are,is
R3726	T14280	T14278	pobj	program,in
R3727	T14281	T14280	amod	embryonic,program
R3728	T14282	T14280	compound	differentiation,program
R3729	T14128	T14129	det	the,functions
R373	T3309	T3307	pobj	genes,by
R3730	T14283	T14284	dep	which,observed
R3731	T14284	T14280	relcl	observed,program
R3732	T14285	T14284	aux	can,observed
R3733	T14129	T14127	nsubj	functions,are
R3734	T14286	T14284	advmod	often,observed
R3735	T14287	T14284	auxpass	be,observed
R3736	T14130	T14129	prep	of,functions
R3737	T14288	T14284	advmod	morphologically,observed
R3738	T14289	T14290	punct	[,38
R3739	T14131	T14132	compound	μ,calpain
R374	T3310	T3309	nmod	Capn1,genes
R3740	T14290	T14284	parataxis	38,observed
R3741	T14291	T14290	punct	],38
R3742	T14292	T14272	punct	.,incurred
R3743	T14132	T14130	pobj	calpain,of
R3744	T14294	T14295	amod	Null,embryos
R3745	T14295	T14296	nsubj	embryos,survive
R3746	T14133	T14132	punct	-,calpain
R3747	T14297	T14295	prep	of,embryos
R3748	T14298	T14299	det	this,type
R3749	T14134	T14127	advmod	simply,are
R375	T3311	T3312	punct	(,k
R3750	T14299	T14297	pobj	type,of
R3751	T14300	T14296	advmod	often,survive
R3752	T14135	T14127	neg	not,are
R3753	T14301	T14296	prep	beyond,survive
R3754	T14302	T14303	det	the,stage
R3755	T14303	T14301	pobj	stage,beyond
R3756	T14136	T14127	acomp	essential,are
R3757	T14304	T14303	compound	morula,stage
R3758	T14305	T14296	punct	", ",survive
R3759	T14306	T14296	cc	and,survive
R376	T3349	T3346	nummod	80,μ
R3760	T14307	T14308	det	a,distribution
R3761	T14308	T14310	nsubjpass	distribution,noted
R3762	T14137	T14090	punct	.,is
R3763	T14309	T14308	amod	Mendelian,distribution
R3764	T14310	T14296	conj	noted,survive
R3765	T14311	T14308	prep	of,distribution
R3766	T14139	T14140	prep	In,demonstrates
R3767	T14312	T14313	amod	embryonic,genotypes
R3768	T14313	T14311	pobj	genotypes,of
R3769	T14314	T14310	auxpass	is,noted
R377	T3312	T3310	parataxis	k,Capn1
R3770	T14315	T14310	advmod	usually,noted
R3771	T14141	T14139	pobj	contrast,In
R3772	T14316	T14310	punct	.,noted
R3773	T14318	T14319	prep	In,thought
R3774	T14142	T14140	punct	", ",demonstrates
R3775	T14320	T14321	amod	other,cases
R3776	T14321	T14318	pobj	cases,In
R3777	T14322	T14319	punct	", ",thought
R3778	T14143	T14140	nsubj	lethality,demonstrates
R3779	T14323	T14319	advmod	however,thought
R378	T3313	T3312	nmod	μ,k
R3780	T14324	T14319	punct	", ",thought
R3781	T14144	T14143	prep	in,lethality
R3782	T14325	T14326	det	the,disruption
R3783	T14326	T14319	nsubjpass	disruption,thought
R3784	T14327	T14326	compound	gene,disruption
R3785	T14145	T14146	nmod	Capn2,embryos
R3786	T14328	T14319	auxpass	is,thought
R3787	T14329	T14330	aux	to,compromise
R3788	T14146	T14144	pobj	embryos,in
R3789	T14330	T14319	xcomp	compromise,thought
R379	T3314	T3312	punct	-,k
R3790	T14331	T14330	advmod	fatally,compromise
R3791	T14332	T14333	det	the,viability
R3792	T14359	T14350	auxpass	are,observed
R3793	T14333	T14330	dobj	viability,compromise
R3794	T14334	T14333	prep	of,viability
R3795	T14335	T14334	pobj	cells,of
R3796	T14336	T14335	prep	in,cells
R3797	T14360	T14350	advmod	ever,observed
R3798	T14337	T14338	det	the,embryo
R3799	T14338	T14336	pobj	embryo,in
R38	T709	T711	nsubjpass	enzymes,composed
R380	T3315	T3312	nummod	80,k
R3800	T14339	T14338	amod	early,embryo
R3801	T14361	T14350	cc	and,observed
R3802	T14340	T14341	punct	[,39
R3803	T14341	T14330	parataxis	39,compromise
R3804	T14342	T14341	dep	3,39
R3805	T14362	T14363	amod	homozygous,cells
R3806	T14343	T14341	punct	",",39
R3807	T14344	T14345	punct	-,43
R3808	T14345	T14341	prep	43,39
R3809	T14363	T14366	nsubjpass	cells,isolated
R381	T3350	T3347	cc	and,k
R3810	T14346	T14341	punct	],39
R3811	T14347	T14319	punct	.,thought
R3812	T14349	T14350	prep	In,observed
R3813	T14364	T14363	compound	mutant,cells
R3814	T14351	T14352	det	these,instances
R3815	T14352	T14349	pobj	instances,In
R3816	T14353	T14350	punct	", ",observed
R3817	T14365	T14363	compound	ES,cells
R3818	T14354	T14355	advmod	only,embryos
R3819	T14355	T14350	nsubjpass	embryos,observed
R382	T3316	T3312	punct	),k
R3820	T14356	T14355	det	a,embryos
R3821	T14366	T14350	conj	isolated,observed
R3822	T14357	T14355	amod	few,embryos
R3823	T14358	T14355	amod	null,embryos
R3824	T14367	T14366	aux	could,isolated
R3825	T14368	T14366	neg	not,isolated
R3826	T14369	T14366	auxpass	be,isolated
R3827	T14370	T14366	punct	.,isolated
R3828	T14372	T14373	det	The,knockout
R3829	T14465	T14467	compound	calpain,involvement
R383	T3317	T3310	cc	and,Capn1
R3830	T14373	T14375	nsubj	knockout,fits
R3831	T14466	T14465	punct	-,calpain
R3832	T14467	T14463	pobj	involvement,of
R3833	T14468	T14467	prep	in,involvement
R3834	T14374	T14373	compound	Capn2,knockout
R3835	T14469	T14470	compound	cell,proliferation
R3836	T14470	T14468	pobj	proliferation,in
R3837	T14376	T14375	prep	into,fits
R3838	T14471	T14467	prep	in,involvement
R3839	T14472	T14473	amod	certain,circumstances
R384	T3318	T3310	conj	Capn2,Capn1
R3840	T14473	T14471	pobj	circumstances,in
R3841	T14474	T14462	punct	", ",reports
R3842	T14377	T14378	det	the,category
R3843	T14475	T14462	prep	including,reports
R3844	T14476	T14475	pobj	reports,including
R3845	T14477	T14476	prep	of,reports
R3846	T14378	T14376	pobj	category,into
R3847	T14478	T14479	poss	its,involvement
R3848	T14479	T14477	pobj	involvement,of
R3849	T14480	T14479	prep	in,involvement
R385	T3351	T3352	nmod	m,k
R3850	T14379	T14378	amod	latter,category
R3851	T14481	T14482	compound	chromosome,segregation
R3852	T14482	T14480	pobj	segregation,in
R3853	T14380	T14375	punct	.,fits
R3854	T14483	T14482	prep	during,segregation
R3855	T14484	T14483	pobj	mitosis,during
R3856	T14485	T14486	punct	[,44
R3857	T14382	T14383	mark	If,is
R3858	T14486	T14484	parataxis	44,mitosis
R3859	T14487	T14486	punct	],44
R386	T3319	T3320	punct	(,k
R3860	T14488	T14489	advmod	as,as
R3861	T14489	T14483	cc	as,during
R3862	T14490	T14489	advmod	well,as
R3863	T14491	T14483	conj	during,during
R3864	T14492	T14491	pobj	meiosis,during
R3865	T14383	T14385	advcl	is,is
R3866	T14493	T14494	punct	[,45
R3867	T14494	T14492	parataxis	45,meiosis
R3868	T14495	T14494	punct	],45
R3869	T14384	T14383	nsubj	it,is
R387	T3320	T3318	parataxis	k,Capn2
R3870	T14496	T14459	punct	.,been
R3871	T14386	T14383	acomp	true,is
R3872	T14498	T14499	nsubj	Defects,contribute
R3873	T14500	T14498	prep	in,Defects
R3874	T14501	T14500	pobj	migration,in
R3875	T14387	T14388	mark	that,is
R3876	T14502	T14498	punct	", ",Defects
R3877	T14503	T14498	acl	reported,Defects
R3878	T14504	T14503	prep	in,reported
R3879	T14388	T14383	ccomp	is,is
R388	T3321	T3320	nmod	m,k
R3880	T14505	T14506	nmod	Capn4,cells
R3881	T14506	T14504	pobj	cells,in
R3882	T14507	T14505	punct	-,Capn4
R3883	T14389	T14390	compound	m,calpain
R3884	T14508	T14505	punct	/,Capn4
R3885	T14509	T14505	punct	-,Capn4
R3886	T14390	T14388	nsubj	calpain,is
R3887	T14510	T14511	punct	[,9
R3888	T14511	T14503	parataxis	9,reported
R3889	T14512	T14511	punct	],9
R389	T3352	T3347	conj	k,k
R3890	T14391	T14390	punct	-,calpain
R3891	T14513	T14499	punct	", ",contribute
R3892	T14514	T14499	aux	could,contribute
R3893	T14515	T14499	prep	to,contribute
R3894	T14392	T14388	acomp	essential,is
R3895	T14516	T14517	amod	failed,development
R3896	T14393	T14392	prep	for,essential
R3897	T14517	T14515	pobj	development,to
R3898	T14518	T14517	amod	embryonic,development
R3899	T14519	T14499	punct	.,contribute
R39	T710	T709	amod	heterodimeric,enzymes
R390	T3322	T3320	punct	-,k
R3900	T14394	T14395	det	some,aspect
R3901	T14521	T14522	det	The,association
R3902	T14522	T14523	nsubjpass	association,noted
R3903	T14395	T14393	pobj	aspect,for
R3904	T14524	T14522	prep	between,association
R3905	T14396	T14395	prep	of,aspect
R3906	T14525	T14524	pobj	calpain,between
R3907	T14526	T14525	cc	and,calpain
R3908	T14397	T14398	compound	cell,viability
R3909	T14527	T14528	compound	cell,viability
R391	T3353	T3351	punct	-,m
R3910	T14528	T14525	conj	viability,calpain
R3911	T14529	T14523	aux	has,noted
R3912	T14398	T14396	pobj	viability,of
R3913	T14530	T14523	auxpass	been,noted
R3914	T14531	T14523	prep	in,noted
R3915	T14532	T14533	nmod	Capn4,MEFs
R3916	T14533	T14531	pobj	MEFs,in
R3917	T14534	T14532	punct	-,Capn4
R3918	T14535	T14532	punct	/,Capn4
R3919	T14536	T14532	punct	-,Capn4
R392	T3354	T3351	nummod	80,m
R3920	T14399	T14385	punct	", ",is
R3921	T14537	T14533	cc	and,MEFs
R3922	T14538	T14539	amod	other,lines
R3923	T14400	T14401	det	the,survival
R3924	T14401	T14385	nsubj	survival,is
R3925	T14539	T14533	conj	lines,MEFs
R3926	T14540	T14539	compound	cell,lines
R3927	T14541	T14523	punct	", ",noted
R3928	T14402	T14401	prep	of,survival
R3929	T14542	T14543	mark	although,distinguish
R393	T3356	T3357	nummod	62,%
R3930	T14543	T14523	advcl	distinguish,noted
R3931	T14403	T14404	det	a,embryos
R3932	T14544	T14545	det	the,work
R3933	T14545	T14543	nsubj	work,distinguish
R3934	T14404	T14402	pobj	embryos,of
R3935	T14546	T14545	amod	reported,work
R3936	T14547	T14543	advmod	frequently,distinguish
R3937	T14405	T14404	amod	few,embryos
R3938	T14548	T14543	aux	did,distinguish
R3939	T14549	T14543	neg	not,distinguish
R394	T3429	T3419	punct	;,levels
R3940	T14550	T14543	prep	between,distinguish
R3941	T14406	T14404	nmod	Capn2,embryos
R3942	T14551	T14552	nmod	μ,calpain
R3943	T14552	T14550	pobj	calpain,between
R3944	T14407	T14406	punct	-,Capn2
R3945	T14553	T14551	punct	-,μ
R3946	T14554	T14551	cc	and,μ
R3947	T14408	T14406	punct	/,Capn2
R3948	T14555	T14551	conj	m,μ
R3949	T14556	T14552	punct	-,calpain
R395	T3357	T3358	compound	%,identity
R3950	T14557	T14543	punct	", ",distinguish
R3951	T14409	T14406	punct	-,Capn2
R3952	T14558	T14543	cc	and,distinguish
R3953	T14559	T14560	aux	did,show
R3954	T14410	T14401	prep	to,survival
R3955	T14560	T14543	conj	show,distinguish
R3956	T14561	T14560	neg	not,show
R3957	T14562	T14563	mark	that,was
R3958	T14411	T14412	det	the,stage
R3959	T14563	T14560	ccomp	was,show
R396	T3430	T3431	quantmod	10,50
R3960	T14564	T14563	nsubj	calpain,was
R3961	T14565	T14566	advmod	strictly,essential
R3962	T14412	T14410	pobj	stage,to
R3963	T14566	T14563	acomp	essential,was
R3964	T14567	T14568	punct	[,6
R3965	T14568	T14560	parataxis	6,show
R3966	T14569	T14568	nummod	5,6
R3967	T14413	T14414	nummod	8,cell
R3968	T14414	T14412	compound	cell,stage
R3969	T14570	T14568	punct	",",6
R397	T3431	T3433	nummod	50,μM
R3970	T14571	T14568	punct	],6
R3971	T14572	T14523	punct	.,noted
R3972	T14415	T14414	punct	-,cell
R3973	T14574	T14575	prep	In,shown
R3974	T14416	T14417	advmod	most,likely
R3975	T14576	T14577	preconj	both,cells
R3976	T14577	T14574	pobj	cells,In
R3977	T14417	T14418	advmod	likely,due
R3978	T14578	T14577	compound	CHO,cells
R3979	T14579	T14577	cc	and,cells
R398	T3358	T3355	dobj	identity,share
R3980	T14580	T14581	nmod	Capn4,MEFs
R3981	T14418	T14385	prep	due,is
R3982	T14581	T14577	conj	MEFs,cells
R3983	T14582	T14580	punct	-,Capn4
R3984	T14583	T14580	punct	/,Capn4
R3985	T14419	T14418	pcomp	to,due
R3986	T14584	T14580	punct	-,Capn4
R3987	T14585	T14575	punct	", ",shown
R3988	T14420	T14421	det	the,persistence
R3989	T14586	T14575	nsubjpass	calpain,shown
R399	T3432	T3431	punct	–,50
R3990	T14587	T14575	auxpass	was,shown
R3991	T14588	T14589	aux	to,influence
R3992	T14421	T14418	pobj	persistence,due
R3993	T14589	T14575	xcomp	influence,shown
R3994	T14590	T14591	compound	cell,proliferation
R3995	T14591	T14589	dobj	proliferation,influence
R3996	T14422	T14421	prep	of,persistence
R3997	T14592	T14589	punct	", ",influence
R3998	T14593	T14594	cc	but,at
R3999	T14594	T14589	prep	at,influence
R4	T666	T669	nsubjpass	Calpain,required
R40	T712	T711	auxpass	are,composed
R400	T3433	T3434	compound	μM,Ca2
R4000	T14423	T14424	det	some,calpain
R4001	T14595	T14594	advmod	only,at
R4002	T14596	T14597	advmod	very,low
R4003	T14597	T14598	amod	low,densities
R4004	T14424	T14422	pobj	calpain,of
R4005	T14598	T14594	pobj	densities,at
R4006	T14599	T14598	compound	cell,densities
R4007	T14600	T14601	punct	[,5
R4008	T14425	T14424	amod	maternal,calpain
R4009	T14601	T14589	parataxis	5,influence
R401	T3434	T3419	appos	Ca2,levels
R4010	T14602	T14601	punct	],5
R4011	T14426	T14424	compound	m,calpain
R4012	T14603	T14575	punct	.,shown
R4013	T14605	T14606	nsubjpass	Calpain,associated
R4014	T14427	T14424	punct	-,calpain
R4015	T14607	T14606	aux	has,associated
R4016	T14428	T14424	appos	mRNA,calpain
R4017	T14608	T14606	advmod	also,associated
R4018	T14609	T14606	auxpass	been,associated
R4019	T14610	T14606	prep	with,associated
R402	T3359	T3360	compound	amino,acid
R4020	T14611	T14610	pobj	progression,with
R4021	T14612	T14611	prep	through,progression
R4022	T14429	T14428	cc	and,mRNA
R4023	T14613	T14614	det	the,stage
R4024	T14614	T14612	pobj	stage,through
R4025	T14615	T14614	compound	G1,stage
R4026	T14430	T14429	punct	/,and
R4027	T14616	T14614	prep	of,stage
R4028	T14617	T14618	det	the,cycle
R4029	T14431	T14429	cc	or,and
R403	T3435	T3434	punct	+,Ca2
R4030	T14618	T14616	pobj	cycle,of
R4031	T14619	T14618	compound	cell,cycle
R4032	T14620	T14621	punct	[,6
R4033	T14432	T14428	conj	protein,mRNA
R4034	T14621	T14606	parataxis	6,associated
R4035	T14622	T14621	punct	],6
R4036	T14623	T14606	punct	.,associated
R4037	T14433	T14421	prep	through,persistence
R4038	T14434	T14435	quantmod	2,3
R4039	T14625	T14626	advmod	Furthermore,reputed
R404	T3436	T3434	prep	for,Ca2
R4040	T14435	T14437	nummod	3,divisions
R4041	T14627	T14626	punct	", ",reputed
R4042	T14436	T14435	punct	–,3
R4043	T14628	T14626	nsubjpass	some,reputed
R4044	T14629	T14628	prep	of,some
R4045	T14630	T14631	det	the,proteins
R4046	T14437	T14433	pobj	divisions,through
R4047	T14631	T14629	pobj	proteins,of
R4048	T14632	T14631	acl	known,proteins
R4049	T14438	T14437	compound	cell,divisions
R405	T3437	T3438	compound	μ,calpain
R4050	T14633	T14634	aux	to,involved
R4051	T14634	T14632	xcomp	involved,known
R4052	T14635	T14634	auxpass	be,involved
R4053	T14439	T14385	punct	.,is
R4054	T14636	T14634	prep	in,involved
R4055	T14637	T14638	compound	cell,cycle
R4056	T14638	T14639	compound	cycle,progression
R4057	T14441	T14442	det	The,reasons
R4058	T14639	T14636	pobj	progression,in
R4059	T14640	T14631	punct	", ",proteins
R406	T3360	T3358	compound	acid,identity
R4060	T14641	T14642	amod	such,as
R4061	T14642	T14631	prep	as,proteins
R4062	T14442	T14444	nsubj	reasons,are
R4063	T14643	T14642	pobj	p53,as
R4064	T14644	T14643	punct	", ",p53
R4065	T14645	T14643	conj	p107,p53
R4066	T14443	T14442	amod	mechanistic,reasons
R4067	T14646	T14645	punct	", ",p107
R4068	T14647	T14648	compound	cyclin,D1
R4069	T14445	T14442	prep	for,reasons
R407	T3438	T3436	pobj	calpain,for
R4070	T14648	T14645	conj	D1,p107
R4071	T14649	T14648	punct	", ",D1
R4072	T14650	T14648	cc	and,D1
R4073	T14651	T14648	conj	p27kip1,D1
R4074	T14652	T14626	auxpass	are,reputed
R4075	T14653	T14654	aux	to,be
R4076	T14446	T14445	pobj	lethality,for
R4077	T14654	T14626	xcomp	be,reputed
R4078	T14655	T14656	compound	calpain,substrates
R4079	T14656	T14654	attr	substrates,be
R408	T3439	T3438	punct	-,calpain
R4080	T14447	T14442	prep	in,reasons
R4081	T14657	T14658	punct	[,3
R4082	T14658	T14654	parataxis	3,be
R4083	T14659	T14658	punct	],3
R4084	T14448	T14449	det	the,absence
R4085	T14660	T14626	punct	.,reputed
R4086	T14662	T14663	compound	Sperm,binding
R4087	T14449	T14447	pobj	absence,in
R4088	T14663	T14664	nsubj	binding,leads
R4089	T14450	T14449	prep	of,absence
R409	T3440	T3434	cc	and,Ca2
R4090	T14665	T14663	prep	to,binding
R4091	T14666	T14667	det	the,oocyte
R4092	T14667	T14665	pobj	oocyte,to
R4093	T14451	T14452	compound	m,calpain
R4094	T14668	T14664	prep	to,leads
R4095	T14669	T14670	amod	increased,calcium
R4096	T14670	T14668	pobj	calcium,to
R4097	T14452	T14450	pobj	calpain,of
R4098	T14671	T14670	amod	cytoplasmic,calcium
R4099	T14672	T14673	dep	which,triggers
R41	T713	T711	prep	of,composed
R410	T3361	T3358	compound	sequence,identity
R4100	T14673	T14670	relcl	triggers,calcium
R4101	T14453	T14452	punct	-,calpain
R4102	T14674	T14675	det	the,reaction
R4103	T14675	T14673	dobj	reaction,triggers
R4104	T14454	T14444	advmod	still,are
R4105	T14455	T14444	acomp	unclear,are
R4106	T14676	T14675	compound	acrosome,reaction
R4107	T14456	T14444	punct	.,are
R4108	T14677	T14678	punct	[,46
R4109	T14678	T14664	parataxis	46,leads
R411	T3441	T3442	quantmod	0.3,0.35
R4110	T14679	T14678	punct	],46
R4111	T14458	T14459	expl	There,been
R4112	T14680	T14664	punct	.,leads
R4113	T14682	T14683	preconj	Both,μ
R4114	T14683	T14684	nmod	μ,calpain
R4115	T14460	T14459	aux	have,been
R4116	T14684	T14689	nsubjpass	calpain,detected
R4117	T14685	T14683	punct	-,μ
R4118	T14686	T14683	cc	and,μ
R4119	T14687	T14683	conj	m,μ
R412	T3442	T3444	nummod	0.35,mM
R4120	T14461	T14462	amod	several,reports
R4121	T14688	T14684	punct	-,calpain
R4122	T14689	T14693	ccomp	detected,implicated
R4123	T14690	T14689	aux	have,detected
R4124	T14691	T14689	advmod	recently,detected
R4125	T14692	T14689	auxpass	been,detected
R4126	T14462	T14459	attr	reports,been
R4127	T14694	T14689	prep	in,detected
R4128	T14695	T14696	compound	rodent,sperm
R4129	T14696	T14694	pobj	sperm,in
R413	T3443	T3442	punct	–,0.35
R4130	T14463	T14462	prep	of,reports
R4131	T14697	T14698	punct	[,47
R4132	T14698	T14696	parataxis	47,sperm
R4133	T14699	T14698	punct	],47
R4134	T14464	T14465	compound	m,calpain
R4135	T14700	T14696	cc	and,sperm
R4136	T14701	T14696	conj	oocytes,sperm
R4137	T14702	T14703	punct	[,45
R4138	T14781	T14782	amod	defective,segregation
R4139	T14703	T14701	parataxis	45,oocytes
R414	T3362	T3355	punct	", ",share
R4140	T14704	T14703	punct	],45
R4141	T14705	T14689	punct	.,detected
R4142	T14706	T14707	compound	m,calpain
R4143	T14782	T14778	conj	segregation,Polyspermy
R4144	T14707	T14693	nsubjpass	calpain,implicated
R4145	T14708	T14707	punct	-,calpain
R4146	T14709	T14693	auxpass	was,implicated
R4147	T14783	T14782	compound	chromosome,segregation
R4148	T14784	T14779	aux	would,have
R4149	T14710	T14693	prep	in,implicated
R415	T3444	T3445	compound	mM,Ca2
R4150	T14711	T14712	det	the,reaction
R4151	T14712	T14710	pobj	reaction,in
R4152	T14785	T14779	advmod	both,have
R4153	T14713	T14712	compound	acrosome,reaction
R4154	T14714	T14715	punct	[,47
R4155	T14786	T14787	amod	lethal,effects
R4156	T14715	T14712	parataxis	47,reaction
R4157	T14716	T14715	punct	],47
R4158	T14717	T14718	dep	which,correlated
R4159	T14718	T14712	relcl	correlated,reaction
R416	T3445	T3434	conj	Ca2,Ca2
R4160	T14787	T14779	dobj	effects,have
R4161	T14719	T14718	prep	with,correlated
R4162	T14720	T14721	det	a,translocation
R4163	T14788	T14787	prep	on,effects
R4164	T14721	T14719	pobj	translocation,with
R4165	T14722	T14721	prep	of,translocation
R4166	T14789	T14790	amod	early,development
R4167	T14723	T14724	compound	m,calpain
R4168	T14724	T14722	pobj	calpain,of
R4169	T14725	T14724	punct	-,calpain
R417	T3363	T3355	cc	and,share
R4170	T14790	T14788	pobj	development,on
R4171	T14726	T14721	prep	to,translocation
R4172	T14727	T14728	det	the,membrane
R4173	T14728	T14726	pobj	membrane,to
R4174	T14729	T14728	amod	cortical,membrane
R4175	T14730	T14728	prep	in,membrane
R4176	T14731	T14730	pobj	oocytes,in
R4177	T14791	T14790	amod	embryonic,development
R4178	T14732	T14733	advmod	where,participate
R4179	T14733	T14728	relcl	participate,membrane
R418	T3446	T3445	punct	+,Ca2
R4180	T14734	T14733	nsubj	it,participate
R4181	T14792	T14779	punct	.,have
R4182	T14735	T14733	aux	might,participate
R4183	T14736	T14733	prep	in,participate
R4184	T14737	T14738	det	the,release
R4185	T14794	T14795	nsubjpass	It,stressed
R4186	T14738	T14736	pobj	release,in
R4187	T14739	T14738	prep	of,release
R4188	T14740	T14741	amod	cortical,contents
R4189	T14741	T14739	pobj	contents,of
R419	T3447	T3445	prep	for,Ca2
R4190	T14796	T14795	aux	should,stressed
R4191	T14742	T14741	compound	granule,contents
R4192	T14743	T14741	acl	required,contents
R4193	T14744	T14745	aux	to,prevent
R4194	T14745	T14743	advcl	prevent,required
R4195	T14797	T14795	advmod	also,stressed
R4196	T14746	T14745	dobj	polyspermy,prevent
R4197	T14747	T14748	punct	[,45
R4198	T14748	T14745	parataxis	45,prevent
R4199	T14749	T14748	punct	],45
R42	T714	T715	amod	distinct,subunits
R420	T3448	T3449	compound	m,calpain
R4200	T14750	T14693	punct	.,implicated
R4201	T14798	T14795	auxpass	be,stressed
R4202	T14752	T14753	compound	m,calpain
R4203	T14753	T14755	nsubj	calpain,relocalized
R4204	T14754	T14753	punct	-,calpain
R4205	T14799	T14800	mark	that,maintained
R4206	T14756	T14755	advmod	also,relocalized
R4207	T14800	T14795	ccomp	maintained,stressed
R4208	T14757	T14755	prep	to,relocalized
R4209	T14758	T14759	det	the,spindle
R421	T3449	T3447	pobj	calpain,for
R4210	T14759	T14757	pobj	spindle,to
R4211	T14801	T14802	nmod	Capn4,cells
R4212	T14760	T14759	nmod	oocyte,spindle
R4213	T14761	T14759	amod	meiotic,spindle
R4214	T14762	T14755	prep	after,relocalized
R4215	T14802	T14800	nsubjpass	cells,maintained
R4216	T14763	T14762	pobj	fertilization,after
R4217	T14764	T14755	punct	", ",relocalized
R4218	T14803	T14801	punct	-,Capn4
R4219	T14765	T14766	dep	were,involved
R422	T3364	T3355	conj	are,share
R4220	T14766	T14755	ccomp	involved,relocalized
R4221	T14767	T14766	nsubjpass	it,involved
R4222	T14804	T14801	punct	/,Capn4
R4223	T14768	T14766	aux	could,involved
R4224	T14769	T14766	auxpass	be,involved
R4225	T14770	T14766	prep	in,involved
R4226	T14771	T14772	compound	chromosome,segregation
R4227	T14772	T14770	pobj	segregation,in
R4228	T14773	T14774	punct	[,45
R4229	T14805	T14801	punct	-,Capn4
R423	T3450	T3449	punct	-,calpain
R4230	T14774	T14766	parataxis	45,involved
R4231	T14775	T14774	punct	],45
R4232	T14776	T14755	punct	.,relocalized
R4233	T14806	T14802	nmod	ES,cells
R4234	T14778	T14779	nsubj	Polyspermy,have
R4235	T14807	T14806	cc	and,ES
R4236	T14780	T14778	cc	or,Polyspermy
R4237	T14808	T14806	conj	MEF,ES
R4238	T14809	T14802	prep	from,cells
R4239	T14810	T14811	det	the,allele
R424	T3451	T3416	punct	.,derive
R4240	T14811	T14809	pobj	allele,from
R4241	T14812	T14811	amod	presumptive,allele
R4242	T14888	T14887	punct	", ",exist
R4243	T14813	T14811	amod	hypomorphic,allele
R4244	T14889	T14890	npadvmod	calpain,independent
R4245	T14814	T14800	aux	can,maintained
R4246	T14890	T14892	amod	independent,mechanisms
R4247	T14891	T14890	punct	-,independent
R4248	T14892	T14887	nsubj	mechanisms,exist
R4249	T14815	T14800	auxpass	be,maintained
R425	T3453	T3454	nsubjpass	It,assumed
R4250	T14893	T14892	prep	for,mechanisms
R4251	T14894	T14893	pcomp	protecting,for
R4252	T14816	T14800	prep	in,maintained
R4253	T14895	T14896	compound	cell,viability
R4254	T14896	T14894	dobj	viability,protecting
R4255	T14817	T14816	pobj	culture,in
R4256	T14897	T14887	aux	might,exist
R4257	T14898	T14887	prep	in,exist
R4258	T14899	T14900	det	these,lines
R4259	T14818	T14800	prep	despite,maintained
R426	T3365	T3366	advmod	very,similar
R4260	T14900	T14898	pobj	lines,in
R4261	T14901	T14900	compound	cell,lines
R4262	T14819	T14820	det	an,lack
R4263	T14902	T14903	dep	that,are
R4264	T14903	T14887	ccomp	are,exist
R4265	T14904	T14903	acomp	absent,are
R4266	T14820	T14818	pobj	lack,despite
R4267	T14905	T14903	prep	in,are
R4268	T14906	T14907	amod	early,cells
R4269	T14821	T14820	amod	apparent,lack
R427	T3455	T3454	auxpass	is,assumed
R4270	T14907	T14905	pobj	cells,in
R4271	T14908	T14907	amod	embryonic,cells
R4272	T14909	T14887	punct	.,exist
R4273	T14822	T14820	prep	of,lack
R4274	T14911	T14912	det	The,failure
R4275	T14912	T14914	nsubj	failure,suggests
R4276	T14823	T14824	compound	calpain,activity
R4277	T14913	T14912	amod	repeated,failure
R4278	T14915	T14916	aux	to,achieve
R4279	T14916	T14912	acl	achieve,failure
R428	T3456	T3454	advmod	generally,assumed
R4280	T14917	T14918	compound	gene,conversion
R4281	T14824	T14822	pobj	activity,of
R4282	T14918	T14916	dobj	conversion,achieve
R4283	T14919	T14918	prep	of,conversion
R4284	T14920	T14919	pobj	Capn2,of
R4285	T14825	T14800	punct	", ",maintained
R4286	T14921	T14920	punct	+,Capn2
R4287	T14922	T14920	punct	/,Capn2
R4288	T14923	T14920	punct	-,Capn2
R4289	T14826	T14827	mark	as,assessed
R429	T3457	T3458	mark	that,maintain
R4290	T14924	T14918	prep	to,conversion
R4291	T14925	T14924	pobj	Capn2,to
R4292	T14827	T14800	advcl	assessed,maintained
R4293	T14926	T14925	punct	-,Capn2
R4294	T14927	T14925	punct	/,Capn2
R4295	T14928	T14925	punct	-,Capn2
R4296	T14828	T14827	prep	by,assessed
R4297	T14929	T14930	compound	ES,cells
R4298	T14930	T14918	appos	cells,conversion
R4299	T14829	T14830	compound	casein,zymography
R43	T715	T713	pobj	subunits,of
R430	T3366	T3364	acomp	similar,are
R4300	T14931	T14918	prep	by,conversion
R4301	T14932	T14931	pobj	selection,by
R4302	T14933	T14932	prep	of,selection
R4303	T14830	T14828	pobj	zymography,by
R4304	T14934	T14933	pobj	clones,of
R4305	T14935	T14932	prep	in,selection
R4306	T14831	T14828	cc	or,by
R4307	T14936	T14937	amod	high,concentrations
R4308	T14937	T14935	pobj	concentrations,in
R4309	T14938	T14937	prep	of,concentrations
R431	T3458	T3454	ccomp	maintain,assumed
R4310	T14832	T14828	conj	by,by
R4311	T14939	T14938	pobj	G418,of
R4312	T14940	T14941	mark	that,compromises
R4313	T14833	T14834	det	the,appearance
R4314	T14941	T14914	ccomp	compromises,suggests
R4315	T14942	T14943	det	the,state
R4316	T14943	T14941	nsubj	state,compromises
R4317	T14834	T14832	pobj	appearance,by
R4318	T14944	T14943	amod	homozygous,state
R4319	T14945	T14943	compound	mutant,state
R432	T3459	T3460	nmod	μ,calpain
R4320	T14946	T14941	advmod	somehow,compromises
R4321	T14835	T14834	prep	of,appearance
R4322	T14947	T14948	compound	cell,viability
R4323	T14948	T14941	dobj	viability,compromises
R4324	T14836	T14837	amod	characteristic,products
R4325	T14949	T14948	cc	or,viability
R4326	T14950	T14951	amod	clonal,selection
R4327	T14951	T14948	conj	selection,viability
R4328	T14837	T14835	pobj	products,of
R4329	T14952	T14951	prep	of,selection
R433	T3367	T3364	prep	in,are
R4330	T14953	T14954	compound	ES,cells
R4331	T14954	T14952	pobj	cells,of
R4332	T14955	T14914	punct	.,suggests
R4333	T14838	T14837	compound	spectrin,products
R4334	T14957	T14958	nsubj	Efforts,are
R4335	T14959	T14960	advmod	under,way
R4336	T14839	T14837	compound	breakdown,products
R4337	T14960	T14958	advmod	way,are
R4338	T14961	T14962	aux	to,express
R4339	T14840	T14795	punct	.,stressed
R434	T3460	T3458	nsubj	calpain,maintain
R4340	T14962	T14958	xcomp	express,are
R4341	T14963	T14964	det	the,subunit
R4342	T14842	T14843	nsubj	It,is
R4343	T14844	T14843	acomp	conceivable,is
R4344	T14964	T14962	dobj	subunit,express
R4345	T14965	T14964	nmod	m,subunit
R4346	T14966	T14965	punct	-,m
R4347	T14845	T14843	punct	", ",is
R4348	T14967	T14965	nummod	80,m
R4349	T14968	T14964	compound	k,subunit
R435	T3461	T3459	punct	-,μ
R4350	T14969	T14964	prep	from,subunit
R4351	T14846	T14847	mark	as,discussed
R4352	T14970	T14971	det	a,transgene
R4353	T14971	T14969	pobj	transgene,from
R4354	T14847	T14843	advcl	discussed,is
R4355	T14972	T14971	compound	Capn2,transgene
R4356	T14973	T14971	compound	cDNA,transgene
R4357	T14848	T14847	advmod	earlier,discussed
R4358	T14974	T14971	compound	rescue,transgene
R4359	T14975	T14976	amod	prior,to
R436	T3462	T3459	cc	and,μ
R4360	T14976	T14962	prep	to,express
R4361	T14849	T14843	punct	", ",is
R4362	T14977	T14978	compound	gene,conversion
R4363	T14978	T14976	pobj	conversion,to
R4364	T14979	T14962	prep	in,express
R4365	T14850	T14851	mark	that,retained
R4366	T14980	T14979	pobj	order,in
R4367	T14981	T14982	aux	to,rescue
R4368	T14851	T14843	ccomp	retained,is
R4369	T14982	T14980	acl	rescue,order
R437	T3463	T3459	conj	m,μ
R4370	T14983	T14982	advmod	preemptively,rescue
R4371	T14984	T14985	compound	m,calpain
R4372	T14852	T14853	det	a,amount
R4373	T14985	T14987	compound	calpain,activity
R4374	T14986	T14985	punct	-,calpain
R4375	T14987	T14982	dobj	activity,rescue
R4376	T14853	T14851	nsubjpass	amount,retained
R4377	T14988	T14989	mark	before,lost
R4378	T14989	T14982	advcl	lost,rescue
R4379	T14990	T14991	det	the,allele
R438	T3368	T3367	pobj	terms,in
R4380	T14854	T14853	compound	trace,amount
R4381	T14991	T14989	nsubjpass	allele,lost
R4382	T14992	T14991	amod	remaining,allele
R4383	T14993	T14991	amod	endogenous,allele
R4384	T14855	T14853	prep	of,amount
R4385	T14856	T14857	compound	calpain,activity
R4386	T14994	T14995	amod	wild,type
R4387	T14995	T14991	compound	type,allele
R4388	T14996	T14995	punct	-,type
R4389	T14857	T14855	pobj	activity,of
R439	T3464	T3460	punct	-,calpain
R4390	T14997	T14989	auxpass	is,lost
R4391	T14998	T14958	punct	.,are
R4392	T14858	T14853	punct	", ",amount
R4393	T15000	T15001	advmod	However,hampered
R4394	T14859	T14853	prep	beneath,amount
R4395	T15002	T15001	punct	", ",hampered
R4396	T15003	T15001	nsubj	difficulty,hampered
R4397	T14860	T14859	pobj	levels,beneath
R4398	T15004	T15003	prep	in,difficulty
R4399	T15005	T15004	pcomp	achieving,in
R44	T716	T715	amod	catalytic,subunits
R440	T3465	T3466	poss	their,sensitivities
R4400	T15006	T15007	amod	stable,expression
R4401	T14861	T14860	amod	detectable,levels
R4402	T15007	T15005	dobj	expression,achieving
R4403	T15008	T15007	prep	of,expression
R4404	T15009	T15010	det	the,transgene
R4405	T14862	T14861	prep	by,detectable
R4406	T15010	T15008	pobj	transgene,of
R4407	T15011	T15010	compound	rescue,transgene
R4408	T14863	T14864	det	these,methods
R4409	T15012	T15001	aux	has,hampered
R441	T3466	T3458	dobj	sensitivities,maintain
R4410	T15013	T15014	advmod	thus,far
R4411	T15014	T15001	advmod	far,hampered
R4412	T14864	T14862	pobj	methods,by
R4413	T15015	T15016	det	these,attempts
R4414	T15016	T15001	dobj	attempts,hampered
R4415	T14865	T14851	punct	", ",retained
R4416	T15017	T15001	punct	.,hampered
R4417	T14866	T14851	auxpass	is,retained
R4418	T15019	T15020	nummod	One,curiosity
R4419	T15020	T15021	nsubj	curiosity,was
R442	T3369	T3368	prep	of,terms
R4420	T14867	T14851	prep	from,retained
R4421	T15022	T15023	dep	which,arose
R4422	T15023	T15020	relcl	arose,curiosity
R4423	T14868	T14869	det	this,allele
R4424	T15024	T15023	prep	during,arose
R4425	T15025	T15026	det	the,genotyping
R4426	T15026	T15024	pobj	genotyping,during
R4427	T14869	T14867	pobj	allele,from
R4428	T15027	T15026	prep	of,genotyping
R4429	T15028	T15027	pobj	progeny,of
R443	T3467	T3466	amod	differential,sensitivities
R4430	T15029	T15028	prep	from,progeny
R4431	T14870	T14869	compound	mutant,allele
R4432	T15030	T15031	amod	heterozygous,interbreeding
R4433	T15031	T15029	pobj	interbreeding,from
R4434	T15032	T15033	det	the,ratio
R4435	T15033	T15021	attr	ratio,was
R4436	T15034	T15035	advmod	highly,non-Mendelian
R4437	T14871	T14869	compound	Capn4,allele
R4438	T15035	T15033	amod	non-Mendelian,ratio
R4439	T15036	T15033	prep	of,ratio
R444	T3468	T3466	prep	to,sensitivities
R4440	T15037	T15036	pobj	Capn2,of
R4441	T14872	T14843	punct	", ",is
R4442	T15038	T15037	punct	+,Capn2
R4443	T15039	T15037	punct	/,Capn2
R4444	T15040	T15037	punct	+,Capn2
R4445	T14873	T14843	cc	and,is
R4446	T15041	T15042	punct	(,%
R4447	T15042	T15037	parataxis	%,Capn2
R4448	T15043	T15042	nummod	11.6,%
R4449	T14874	T14875	nsubj	it,is
R445	T3370	T3369	pobj	structure,of
R4450	T15044	T15042	punct	),%
R4451	T15045	T15033	prep	to,ratio
R4452	T14875	T14843	conj	is,is
R4453	T15046	T15045	pobj	Capn2,to
R4454	T15047	T15046	punct	+,Capn2
R4455	T15048	T15046	punct	/,Capn2
R4456	T14876	T14875	acomp	sufficient,is
R4457	T15049	T15046	punct	-,Capn2
R4458	T14877	T14876	prep	for,sufficient
R4459	T15050	T15051	punct	(,%
R446	T3469	T3468	pobj	calcium,to
R4460	T14878	T14877	pcomp	maintaining,for
R4461	T15051	T15046	parataxis	%,Capn2
R4462	T15052	T15051	nummod	88.4,%
R4463	T14879	T14880	det	the,viability
R4464	T15053	T15051	punct	),%
R4465	T15054	T15033	prep	of,ratio
R4466	T15055	T15054	pobj	weanlings,of
R4467	T14880	T14878	dobj	viability,maintaining
R4468	T15056	T15021	punct	.,was
R4469	T15058	T15059	prep	At,is
R447	T3470	T3471	advmod	in,vivo
R4470	T14881	T14880	prep	of,viability
R4471	T15060	T15058	amod	present,At
R4472	T14882	T14883	det	the,cells
R4473	T15061	T15059	punct	", ",is
R4474	T15062	T15059	expl	there,is
R4475	T14883	T14881	pobj	cells,of
R4476	T15063	T15064	det	no,explanation
R4477	T15064	T15059	attr	explanation,is
R4478	T15065	T15064	amod	obvious,explanation
R4479	T14884	T14875	punct	.,is
R448	T3471	T3458	advmod	vivo,maintain
R4480	T15066	T15064	prep	for,explanation
R4481	T15067	T15068	det	this,result
R4482	T14886	T14887	advmod	Furthermore,exist
R4483	T15068	T15066	pobj	result,for
R4484	T15069	T15059	punct	.,is
R4485	T15071	T15072	nsubj	Crosses,produced
R4486	T15073	T15071	prep	between,Crosses
R4487	T15074	T15075	amod	wild,type
R4488	T15100	T15098	pobj	line,involving
R4489	T15075	T15077	nmod	type,mice
R449	T3371	T3370	punct	", ",structure
R4490	T15076	T15075	punct	-,type
R4491	T15077	T15073	pobj	mice,between
R4492	T15078	T15075	cc	and,type
R4493	T15079	T15075	conj	heterozygous,type
R4494	T15080	T15072	advmod	also,produced
R4495	T15101	T15100	nmod	Capn4,line
R4496	T15081	T15072	dobj	progeny,produced
R4497	T15082	T15081	prep	with,progeny
R4498	T15083	T15084	det	a,proportion
R4499	T15084	T15082	pobj	proportion,with
R45	T717	T715	punct	", ",subunits
R450	T3472	T3458	punct	", ",maintain
R4500	T15102	T15100	amod	transgenic,line
R4501	T15085	T15084	amod	greater,proportion
R4502	T15103	T15100	acl	generated,line
R4503	T15086	T15087	mark	than,expected
R4504	T15087	T15085	advcl	expected,greater
R4505	T15104	T15103	agent	by,generated
R4506	T15088	T15084	prep	of,proportion
R4507	T15089	T15090	amod	heterozygous,offspring
R4508	T15105	T15104	pobj	Zimmerman,by
R4509	T15090	T15088	pobj	offspring,of
R451	T3473	T3474	mark	although,demonstrated
R4510	T15091	T15072	punct	.,produced
R4511	T15106	T15105	cc	and,Zimmerman
R4512	T15093	T15094	advmod	Interestingly,yielded
R4513	T15107	T15105	conj	colleagues,Zimmerman
R4514	T15095	T15094	punct	", ",yielded
R4515	T15096	T15097	amod	heterozygous,crosses
R4516	T15097	T15094	nsubj	crosses,yielded
R4517	T15108	T15094	dobj	progeny,yielded
R4518	T15098	T15097	prep	involving,crosses
R4519	T15099	T15100	det	the,line
R452	T3372	T3373	compound	protein,chemistry
R4520	T15109	T15108	prep	with,progeny
R4521	T15110	T15111	det	a,skewing
R4522	T15111	T15109	pobj	skewing,with
R4523	T15112	T15111	amod	similar,skewing
R4524	T15113	T15114	punct	", ",extreme
R4525	T15114	T15111	parataxis	extreme,skewing
R4526	T15115	T15114	mark	if,extreme
R4527	T15116	T15114	advmod	less,extreme
R4528	T15117	T15114	punct	", ",extreme
R4529	T15118	T15111	prep	in,skewing
R453	T3373	T3370	conj	chemistry,structure
R4530	T15119	T15118	pobj	favor,in
R4531	T15120	T15119	prep	of,favor
R4532	T15121	T15122	det	the,genotype
R4533	T15122	T15120	pobj	genotype,of
R4534	T15123	T15122	amod	heterozygous,genotype
R4535	T15124	T15094	punct	.,yielded
R4536	T15126	T15127	prep	Out,were
R4537	T15128	T15126	prep	of,Out
R4538	T15129	T15130	det	a,total
R4539	T15130	T15128	pobj	total,of
R454	T3474	T3458	advcl	demonstrated,maintain
R4540	T15131	T15130	prep	of,total
R4541	T15132	T15133	nummod	80,animals
R4542	T15133	T15131	pobj	animals,of
R4543	T15134	T15133	amod	genotyped,animals
R4544	T15135	T15127	punct	", ",were
R4545	T15136	T15137	nummod	22.5,%
R4546	T15137	T15127	nsubj	%,were
R4547	T15138	T15139	amod	wild,type
R4548	T15139	T15127	attr	type,were
R4549	T15140	T15139	punct	-,type
R455	T3475	T3474	nsubjpass	this,demonstrated
R4550	T15141	T15127	cc	and,were
R4551	T15142	T15143	nummod	77.5,%
R4552	T15143	T15144	nsubj	%,were
R4553	T15144	T15127	conj	were,were
R4554	T15145	T15144	acomp	heterozygous,were
R4555	T15146	T15127	punct	", ",were
R4556	T15147	T15148	mark	with,observed
R4557	T15148	T15127	advcl	observed,were
R4558	T15149	T15150	det	no,progeny
R4559	T15150	T15148	nsubj	progeny,observed
R456	T3476	T3474	aux	has,demonstrated
R4560	T15151	T15152	amod	homozygous,null
R4561	T15152	T15150	amod	null,progeny
R4562	T15153	T15154	punct	[,32
R4563	T15154	T15148	parataxis	32,observed
R4564	T15155	T15154	punct	],32
R4565	T15156	T15127	punct	.,were
R4566	T15158	T15159	det	These,observations
R4567	T15159	T15160	nsubj	observations,suggest
R4568	T15161	T15162	det	a,advantage
R4569	T15162	T15160	dobj	advantage,suggest
R457	T3374	T3373	punct	", ",chemistry
R4570	T15163	T15162	amod	developmental,advantage
R4571	T15164	T15162	acl	associated,advantage
R4572	T15165	T15164	prep	with,associated
R4573	T15166	T15167	amod	reduced,expression
R4574	T15167	T15165	pobj	expression,with
R4575	T15168	T15167	compound	calpain,expression
R4576	T15169	T15160	punct	.,suggest
R4577	T15171	T15172	advmod	Perhaps,reveal
R4578	T15173	T15174	amod	future,studies
R4579	T15174	T15172	nsubj	studies,reveal
R458	T3477	T3474	neg	not,demonstrated
R4580	T15175	T15172	aux	will,reveal
R4581	T15176	T15177	det	a,basis
R4582	T15177	T15172	dobj	basis,reveal
R4583	T15178	T15177	amod	mechanistic,basis
R4584	T15179	T15177	prep	for,basis
R4585	T15180	T15179	pobj	this,for
R4586	T15181	T15172	punct	.,reveal
R4589	T15430	T15431	det	The,work
R459	T3478	T3474	advmod	yet,demonstrated
R4590	T15431	T15432	nsubj	work,clarified
R4591	T15433	T15431	acl	presented,work
R4592	T15434	T15433	advmod	here,presented
R4593	T15435	T15432	aux	has,clarified
R4594	T15436	T15437	nummod	two,questions
R4595	T15437	T15432	dobj	questions,clarified
R4596	T15438	T15437	amod	important,questions
R4597	T15439	T15437	prep	regarding,questions
R4598	T15440	T15441	det	the,roles
R4599	T15441	T15439	pobj	roles,regarding
R46	T718	T715	acl	encoded,subunits
R460	T3375	T3373	cc	and,chemistry
R4600	T15442	T15441	amod	physiological,roles
R4601	T15443	T15441	prep	of,roles
R4602	T15444	T15445	det	the,isoforms
R4603	T15445	T15443	pobj	isoforms,of
R4604	T15446	T15445	nummod	two,isoforms
R4605	T15447	T15445	amod	ubiquitous,isoforms
R4606	T15448	T15445	compound	calpain,isoforms
R4607	T15449	T15445	punct	", ",isoforms
R4608	T15450	T15451	nmod	μ,calpain
R4609	T15451	T15445	appos	calpain,isoforms
R461	T3479	T3474	auxpass	been,demonstrated
R4610	T15452	T15450	punct	-,μ
R4611	T15453	T15450	cc	and,μ
R4612	T15454	T15450	conj	m,μ
R4613	T15455	T15451	punct	-,calpain
R4614	T15456	T15432	punct	.,clarified
R4615	T15458	T15459	advmod	Firstly,determined
R4616	T15460	T15459	punct	", ",determined
R4617	T15461	T15459	nsubjpass	it,determined
R4618	T15462	T15459	auxpass	was,determined
R4619	T15463	T15464	mark	that,plays
R462	T3480	T3474	advmod	strictly,demonstrated
R4620	T15464	T15459	ccomp	plays,determined
R4621	T15465	T15466	compound	m,calpain
R4622	T15466	T15464	nsubj	calpain,plays
R4623	T15467	T15466	punct	-,calpain
R4624	T15468	T15469	det	an,role
R4625	T15469	T15464	dobj	role,plays
R4626	T15470	T15469	amod	indispensable,role
R4627	T15471	T15469	prep	in,role
R4628	T15472	T15473	amod	murine,embryogenesis
R4629	T15473	T15471	pobj	embryogenesis,in
R463	T3481	T3454	punct	.,assumed
R4630	T15474	T15469	punct	", ",role
R4631	T15475	T15476	advmod	possibly,related
R4632	T15476	T15469	acl	related,role
R4633	T15477	T15476	prep	to,related
R4634	T15478	T15479	amod	pre-implantation,development
R4635	T15479	T15477	pobj	development,to
R4636	T15480	T15459	punct	.,determined
R4637	T15482	T15483	advmod	Furthermore,carried
R4638	T15484	T15483	punct	", ",carried
R4639	T15485	T15486	det	this,function
R464	T3376	T3377	advmod	in,vitro
R4640	T15486	T15483	nsubjpass	function,carried
R4641	T15487	T15483	aux	can,carried
R4642	T15488	T15483	neg	not,carried
R4643	T15489	T15483	auxpass	be,carried
R4644	T15490	T15483	prt	out,carried
R4645	T15491	T15483	agent	by,carried
R4646	T15492	T15493	compound	μ,calpain
R4647	T15493	T15491	pobj	calpain,by
R4648	T15494	T15493	punct	-,calpain
R4649	T15495	T15483	prep	despite,carried
R465	T3483	T3484	advmod	Furthermore,assumed
R4650	T15496	T15497	det	the,similarities
R4651	T15497	T15495	pobj	similarities,despite
R4652	T15498	T15497	amod	apparent,similarities
R4653	T15499	T15500	advmod	in,vitro
R4654	T15500	T15497	amod	vitro,similarities
R4655	T15501	T15497	prep	of,similarities
R4656	T15502	T15503	nmod	μ,calpain
R4657	T15503	T15501	pobj	calpain,of
R4658	T15504	T15502	punct	-,μ
R4659	T15505	T15502	cc	and,μ
R466	T3377	T3378	amod	vitro,specificity
R4660	T15506	T15502	conj	m,μ
R4661	T15507	T15503	punct	-,calpain
R4662	T15508	T15483	punct	", ",carried
R4663	T15509	T15483	advcl	demonstrating,carried
R4664	T15510	T15511	mark	that,have
R4665	T15511	T15509	ccomp	have,demonstrating
R4666	T15512	T15513	det	the,isoforms
R4667	T15513	T15511	nsubj	isoforms,have
R4668	T15514	T15513	nummod	two,isoforms
R4669	T15515	T15511	advmod	clearly,have
R467	T3485	T3484	punct	", ",assumed
R4670	T15516	T15517	det	some,roles
R4671	T15517	T15511	dobj	roles,have
R4672	T15518	T15517	amod	distinct,roles
R4673	T15519	T15520	advmod	in,vivo
R4674	T15520	T15511	advmod	vivo,have
R4675	T15521	T15483	punct	.,carried
R4676	T15523	T15524	det	The,functions
R4677	T15524	T15525	nsubj	functions,remain
R4678	T15526	T15524	prep	of,functions
R4679	T15527	T15528	compound	m,calpain
R468	T3486	T3487	mark	since,is
R4680	T15528	T15526	pobj	calpain,of
R4681	T15529	T15528	punct	-,calpain
R4682	T15530	T15524	prep	during,functions
R4683	T15531	T15532	amod	post-implantation,embryogenesis
R4684	T15532	T15530	pobj	embryogenesis,during
R4685	T15533	T15530	cc	and,during
R4686	T15534	T15530	conj	in,during
R4687	T15535	T15536	amod	adult,mice
R4688	T15536	T15534	pobj	mice,in
R4689	T15537	T15538	aux	to,elucidated
R469	T3378	T3373	conj	specificity,chemistry
R4690	T15538	T15525	xcomp	elucidated,remain
R4691	T15539	T15538	auxpass	be,elucidated
R4692	T15540	T15525	cc	and,remain
R4693	T15541	T15542	aux	will,have
R4694	T15542	T15525	conj	have,remain
R4695	T15543	T15544	aux	to,addressed
R4696	T15544	T15542	xcomp	addressed,have
R4697	T15545	T15544	auxpass	be,addressed
R4698	T15546	T15544	advcl	using,addressed
R4699	T15547	T15548	det	a,strategy
R47	T719	T718	agent	by,encoded
R470	T3487	T3484	advcl	is,assumed
R4700	T15548	T15546	dobj	strategy,using
R4701	T15549	T15548	amod	conditional,strategy
R4702	T15550	T15551	compound	gene,targeting
R4703	T15551	T15548	compound	targeting,strategy
R4704	T15552	T15525	punct	.,remain
R4705	T16009	T16008	cc	and,Cloning
R4706	T16010	T16008	conj	sequencing,Cloning
R4707	T16011	T16008	prep	of,Cloning
R4708	T16012	T16013	det	the,locus
R4709	T16013	T16011	pobj	locus,of
R471	T3488	T3489	det	the,concentration
R4710	T16014	T16013	compound	mouse,locus
R4711	T16015	T16013	compound	Capn2,locus
R4712	T16017	T16018	det	The,gene
R4713	T16018	T16021	nsubj	gene,encodes
R4714	T16019	T16018	compound	mouse,gene
R4715	T16020	T16018	compound	Capn2,gene
R4716	T16022	T16023	det	the,acids
R4717	T16023	T16021	dobj	acids,encodes
R4718	T16024	T16023	nummod	700,acids
R4719	T16025	T16023	compound	amino,acids
R472	T3379	T3378	compound	substrate,specificity
R4720	T16026	T16023	prep	of,acids
R4721	T16027	T16028	det	the,subunit
R4722	T16028	T16026	pobj	subunit,of
R4723	T16029	T16030	nmod	m,calpain
R4724	T16030	T16028	nmod	calpain,subunit
R4725	T16031	T16030	punct	-,calpain
R4726	T16032	T16028	amod	large,subunit
R4727	T16033	T16034	punct	(,k
R4728	T16034	T16028	parataxis	k,subunit
R4729	T16035	T16034	nmod	m,k
R473	T3489	T3487	nsubj	concentration,is
R4730	T16036	T16034	punct	-,k
R4731	T16037	T16034	nummod	80,k
R4732	T16038	T16034	punct	),k
R4733	T16039	T16021	cc	and,encodes
R4734	T16040	T16021	conj	consists,encodes
R4735	T16041	T16040	prep	of,consists
R4736	T16042	T16043	nummod	21,exons
R4737	T16043	T16041	pobj	exons,of
R4738	T16044	T16043	acl	extending,exons
R4739	T16045	T16044	prep	over,extending
R474	T3490	T3491	amod	cytoplasmic,free
R4740	T16046	T16047	nummod	50,kb
R4741	T16047	T16045	pobj	kb,over
R4742	T16048	T16047	punct	-,kb
R4743	T16049	T16044	prep	on,extending
R4744	T16050	T16049	pobj	chromosome,on
R4745	T16051	T16050	nummod	1,chromosome
R4746	T16052	T16021	punct	.,encodes
R4747	T16054	T16055	det	A,clone
R4748	T16055	T16057	nsubjpass	clone,purchased
R4749	T16056	T16055	compound	cDNA,clone
R475	T3491	T3489	amod	free,concentration
R4750	T16058	T16055	acl	encoding,clone
R4751	T16059	T16060	det	a,portion
R4752	T16060	T16058	dobj	portion,encoding
R4753	T16061	T16060	prep	of,portion
R4754	T16062	T16063	det	the,subunit
R4755	T16063	T16061	pobj	subunit,of
R4756	T16064	T16063	nmod	mouse,subunit
R4757	T16065	T16066	nmod	m,k
R4758	T16066	T16063	compound	k,subunit
R4759	T16067	T16066	punct	-,k
R476	T3380	T3355	punct	.,share
R4760	T16068	T16066	nummod	80,k
R4761	T16069	T16057	auxpass	was,purchased
R4762	T16070	T16071	punct	(,LLNL
R4763	T16071	T16057	parataxis	LLNL,purchased
R4764	T16072	T16073	compound	dbEST,Id
R4765	T16073	T16071	dep	Id,LLNL
R4766	T16074	T16071	punct	", ",LLNL
R4767	T16075	T16071	dep	807416,LLNL
R4768	T16076	T16071	punct	", ",LLNL
R4769	T16077	T16071	dep	Image,LLNL
R477	T3492	T3489	nmod	Ca2,concentration
R4770	T16078	T16071	punct	:,LLNL
R4771	T16079	T16071	dep	606689,LLNL
R4772	T16080	T16071	punct	", ",LLNL
R4773	T16081	T16082	compound	Image,Consortium
R4774	T16082	T16071	dep	Consortium,LLNL
R4775	T16083	T16071	punct	", ",LLNL
R4776	T16084	T16071	punct	),LLNL
R4777	T16085	T16057	punct	.,purchased
R4778	T16087	T16088	nsubjpass	It,found
R4779	T16089	T16088	auxpass	was,found
R478	T3493	T3492	punct	+,Ca2
R4780	T16090	T16091	aux	to,contain
R4781	T16091	T16088	xcomp	contain,found
R4782	T16092	T16093	nummod	2.8,kb
R4783	T16093	T16091	dobj	kb,contain
R4784	T16094	T16093	punct	-,kb
R4785	T16095	T16093	prep	of,kb
R4786	T16096	T16095	pobj	sequence,of
R4787	T16097	T16096	prep	from,sequence
R4788	T16098	T16097	pobj	position,from
R4789	T16099	T16098	nummod	247,position
R479	T3494	T3487	advmod	typically,is
R4790	T16100	T16098	prep	of,position
R4791	T16101	T16102	det	the,sequence
R4792	T16102	T16100	pobj	sequence,of
R4793	T16103	T16102	compound	coding,sequence
R4794	T16104	T16097	prep	to,from
R4795	T16105	T16106	det	the,codon
R4796	T16106	T16104	pobj	codon,to
R4797	T16107	T16106	compound	stop,codon
R4798	T16108	T16106	prep	at,codon
R4799	T16109	T16108	pobj	position,at
R48	T720	T719	pobj	Capn1,by
R480	T3495	T3496	amod	less,1
R4800	T16110	T16109	nummod	"2,101",position
R4801	T16111	T16096	punct	", ",sequence
R4802	T16112	T16096	prep	including,sequence
R4803	T16113	T16114	det	the,UTR
R4804	T16114	T16112	pobj	UTR,including
R4805	T16115	T16114	nummod	3,UTR
R4806	T16116	T16115	punct	',3
R4807	T16117	T16114	punct	-,UTR
R4808	T16118	T16114	prep	to,UTR
R4809	T16119	T16120	det	the,signal
R481	T3496	T3498	nummod	1,μM
R4810	T16120	T16118	pobj	signal,to
R4811	T16121	T16120	compound	polyA,signal
R4812	T16122	T16088	punct	.,found
R4813	T16124	T16125	det	A,fragment
R4814	T16125	T16126	nsubjpass	fragment,subcloned
R4815	T16127	T16125	prep	of,fragment
R4816	T16128	T16129	det	this,cDNA
R4817	T16129	T16127	pobj	cDNA,of
R4818	T16130	T16126	auxpass	was,subcloned
R4819	T16131	T16126	cc	and,subcloned
R482	T3497	T3496	quantmod	than,1
R4820	T16132	T16126	conj	used,subcloned
R4821	T16133	T16134	aux	to,screen
R4822	T16134	T16132	advcl	screen,used
R4823	T16135	T16136	det	a,library
R4824	T16136	T16134	dobj	library,screen
R4825	T16137	T16136	nummod	129Sv,library
R4826	T16138	T16136	nmod	mouse,library
R4827	T16139	T16136	amod	genomic,library
R4828	T16140	T16134	prep	in,screen
R4829	T16141	T16142	compound	λ,Dash
R483	T3382	T3383	prep	Despite,suggest
R4830	T16142	T16140	pobj	Dash,in
R4831	T16143	T16142	punct	-,Dash
R4832	T16144	T16142	nummod	II,Dash
R4833	T16145	T16126	punct	.,subcloned
R4834	T16147	T16148	amod	Overlapping,clones
R4835	T16148	T16150	nsubjpass	clones,obtained
R4836	T16149	T16148	amod	genomic,clones
R4837	T16151	T16150	auxpass	were,obtained
R4838	T16152	T16150	advcl	covering,obtained
R4839	T16153	T16154	nummod	"15,354",bp
R484	T3498	T3487	attr	μM,is
R4840	T16154	T16152	dobj	bp,covering
R4841	T16155	T16150	punct	", ",obtained
R4842	T16156	T16150	advcl	extending,obtained
R4843	T16157	T16156	prep	from,extending
R4844	T16158	T16159	det	a,site
R4845	T16159	T16157	pobj	site,from
R4846	T16160	T16159	compound	BamHI,site
R4847	T16161	T16159	prep	in,site
R4848	T16162	T16161	pobj	intron,in
R4849	T16163	T16162	nummod	3,intron
R485	T3499	T3484	punct	", ",assumed
R4850	T16164	T16157	prep	to,from
R4851	T16165	T16166	det	a,site
R4852	T16166	T16164	pobj	site,to
R4853	T16167	T16166	compound	BamHI,site
R4854	T16168	T16166	prep	in,site
R4855	T16169	T16168	pobj	intron,in
R4856	T16170	T16169	nummod	15,intron
R4857	T16171	T16150	punct	.,obtained
R4858	T16173	T16174	det	These,clones
R4859	T16174	T16175	nsubjpass	clones,sequenced
R486	T3500	T3484	nsubjpass	it,assumed
R4860	T16176	T16175	auxpass	were,sequenced
R4861	T16177	T16175	advmod	completely,sequenced
R4862	T16178	T16175	prep	on,sequenced
R4863	T16179	T16180	det	both,strands
R4864	T16180	T16178	pobj	strands,on
R4865	T16181	T16175	cc	and,sequenced
R4866	T16182	T16183	det	the,data
R4867	T16183	T16184	nsubjpass	data,submitted
R4868	T16184	T16175	conj	submitted,sequenced
R4869	T16185	T16184	auxpass	were,submitted
R487	T3501	T3484	auxpass	is,assumed
R4870	T16186	T16184	prep	to,submitted
R4871	T16187	T16186	pobj	GenBank,to
R4872	T16188	T16189	punct	(,AF497625
R4873	T16189	T16184	parataxis	AF497625,submitted
R4874	T16190	T16189	compound	accession,AF497625
R4875	T16191	T16189	compound	no.,AF497625
R4876	T16192	T16189	punct	),AF497625
R4877	T16193	T16184	punct	.,submitted
R4878	T16195	T16196	det	The,sequences
R4879	T16196	T16198	nsubj	sequences,agreed
R488	T3384	T3385	det	these,similarities
R4880	T16197	T16196	amod	submitted,sequences
R4881	T16199	T16198	advmod	precisely,agreed
R4882	T16200	T16198	prep	with,agreed
R4883	T16201	T16202	det	the,databases
R4884	T16202	T16200	pobj	databases,with
R4885	T16203	T16202	amod	public,databases
R4886	T16204	T16198	punct	", ",agreed
R4887	T16205	T16198	cc	and,agreed
R4888	T16206	T16207	advmod	also,filled
R4889	T16207	T16198	conj	filled,agreed
R489	T3502	T3484	advmod	also,assumed
R4890	T16208	T16207	prt	in,filled
R4891	T16209	T16210	amod	several,gaps
R4892	T16210	T16207	dobj	gaps,filled
R4893	T16211	T16210	amod	small,gaps
R4894	T16212	T16210	acl	corresponding,gaps
R4895	T16213	T16212	prep	to,corresponding
R4896	T16214	T16215	amod	short,sequences
R4897	T16215	T16213	pobj	sequences,to
R4898	T16216	T16215	amod	repetitive,sequences
R4899	T16217	T16218	dep	which,established
R49	T721	T720	punct	(,Capn1
R490	T3503	T3504	mark	that,contribute
R4900	T16218	T16215	relcl	established,sequences
R4901	T16219	T16218	aux	could,established
R4902	T16220	T16218	advmod	only,established
R4903	T16221	T16218	auxpass	be,established
R4904	T16222	T16218	advmod	firmly,established
R4905	T16223	T16218	prep	by,established
R4906	T16224	T16225	amod	repeated,sequencing
R4907	T16225	T16223	pobj	sequencing,by
R4908	T16226	T16225	prep	in,sequencing
R4909	T16227	T16228	amod	non-standard,conditions
R491	T3504	T3484	ccomp	contribute,assumed
R4910	T16228	T16226	pobj	conditions,in
R4911	T16229	T16198	punct	.,agreed
R4912	T16710	T16709	prep	of,Construction
R4913	T16711	T16712	det	the,vector
R4914	T16712	T16710	pobj	vector,of
R4915	T16713	T16712	compound	Capn2,vector
R4916	T16714	T16712	compound	targeting,vector
R4917	T16716	T16717	det	A,construct
R4918	T16717	T16719	nsubjpass	construct,designed
R4919	T16718	T16717	compound	targeting,construct
R492	T3505	T3506	amod	other,factors
R4920	T16720	T16719	auxpass	was,designed
R4921	T16721	T16722	aux	to,replace
R4922	T16722	T16719	advcl	replace,designed
R4923	T16723	T16724	det	a,fragment
R4924	T16724	T16722	dobj	fragment,replace
R4925	T16725	T16726	nummod	785,bp
R4926	T16726	T16724	compound	bp,fragment
R4927	T16727	T16728	compound	BamHI,HindIII
R4928	T16728	T16724	compound	HindIII,fragment
R4929	T16729	T16728	punct	-,HindIII
R493	T3506	T3504	nsubj	factors,contribute
R4930	T16730	T16724	punct	", ",fragment
R4931	T16731	T16724	acl	containing,fragment
R4932	T16732	T16731	dobj	exon,containing
R4933	T16733	T16732	nummod	7,exon
R4934	T16734	T16732	prep	of,exon
R4935	T16735	T16736	det	the,gene
R4936	T16736	T16734	pobj	gene,of
R4937	T16737	T16736	compound	Capn2,gene
R4938	T16738	T16722	punct	", ",replace
R4939	T16739	T16722	prep	with,replace
R494	T3385	T3382	pobj	similarities,Despite
R4940	T16740	T16741	det	the,cassette
R4941	T16741	T16739	pobj	cassette,with
R4942	T16742	T16743	compound	PGK,Neo
R4943	T16743	T16741	compound	Neo,cassette
R4944	T16744	T16743	punct	-,Neo
R4945	T16745	T16746	punct	(,Figure
R4946	T16746	T16722	parataxis	Figure,replace
R4947	T16747	T16746	nummod	1,Figure
R4948	T16748	T16746	punct	),Figure
R4949	T16749	T16719	punct	.,designed
R495	T3507	T3508	advmod	in,vivo
R4950	T16751	T16752	nsubj	Exon,encodes
R4951	T16753	T16751	nummod	7,Exon
R4952	T16754	T16755	nummod	24,acids
R4953	T16755	T16752	dobj	acids,encodes
R4954	T16756	T16755	compound	amino,acids
R4955	T16757	T16752	prep	in,encodes
R4956	T16758	T16759	det	the,region
R4957	T16759	T16757	pobj	region,in
R4958	T16760	T16761	amod	active,site
R4959	T16761	T16759	compound	site,region
R496	T3508	T3506	amod	vivo,factors
R4960	T16762	T16761	punct	-,site
R4961	T16763	T16752	punct	", ",encodes
R4962	T16764	T16752	prep	including,encodes
R4963	T16765	T16764	pobj	Asn286,including
R4964	T16766	T16765	punct	", ",Asn286
R4965	T16767	T16765	appos	one,Asn286
R4966	T16768	T16767	prep	of,one
R4967	T16769	T16770	det	the,residues
R4968	T16770	T16768	pobj	residues,of
R4969	T16771	T16770	amod	catalytic,residues
R497	T3509	T3504	aux	must,contribute
R4970	T16772	T16770	compound	triad,residues
R4971	T16773	T16752	punct	.,encodes
R4972	T16775	T16776	det	The,arm
R4973	T16776	T16781	nsubjpass	arm,provided
R4974	T16777	T16776	amod	short,arm
R4975	T16778	T16776	punct	(,arm
R4976	T16779	T16776	amod	upstream,arm
R4977	T16780	T16776	punct	),arm
R4978	T16782	T16776	prep	of,arm
R4979	T16783	T16784	det	the,construct
R498	T3510	T3504	prep	to,contribute
R4980	T16784	T16782	pobj	construct,of
R4981	T16785	T16784	compound	targeting,construct
R4982	T16786	T16781	auxpass	was,provided
R4983	T16787	T16781	agent	by,provided
R4984	T16788	T16789	det	a,fragment
R4985	T16789	T16787	pobj	fragment,by
R4986	T16790	T16791	nummod	2.7,kb
R4987	T16791	T16789	compound	kb,fragment
R4988	T16792	T16791	punct	-,kb
R4989	T16793	T16794	compound	HindIII,BamHI
R499	T3386	T3383	punct	", ",suggest
R4990	T16794	T16789	compound	BamHI,fragment
R4991	T16795	T16794	punct	-,BamHI
R4992	T16796	T16789	punct	", ",fragment
R4993	T16797	T16789	acl	containing,fragment
R4994	T16798	T16799	nmod	exons,5
R4995	T16799	T16797	dobj	5,containing
R4996	T16800	T16799	cc	and,5
R4997	T16801	T16799	conj	6,5
R4998	T16802	T16789	punct	", ",fragment
R4999	T16803	T16804	dep	which,inserted
R5	T668	T666	punct	-,Calpain
R50	T722	T723	compound	μ,calpain
R500	T3511	T3510	pobj	regulation,to
R5000	T16804	T16789	relcl	inserted,fragment
R5001	T16805	T16804	auxpass	was,inserted
R5002	T16806	T16804	prep	into,inserted
R5003	T16807	T16808	det	the,vector
R5004	T16808	T16806	pobj	vector,into
R5005	T16809	T16808	compound	pPNT,vector
R5006	T16810	T16804	advmod	upstream,inserted
R5007	T16811	T16810	prep	of,upstream
R5008	T16812	T16813	det	the,cassette
R5009	T16813	T16811	pobj	cassette,of
R501	T3512	T3511	prep	of,regulation
R5010	T16814	T16815	compound	PGK,Neo
R5011	T16815	T16813	compound	Neo,cassette
R5012	T16816	T16815	punct	-,Neo
R5013	T16817	T16818	punct	[,48
R5014	T16818	T16804	parataxis	48,inserted
R5015	T16819	T16818	punct	],48
R5016	T16820	T16781	punct	.,provided
R5017	T16822	T16823	prep	During,abolished
R5018	T16824	T16822	pobj	cloning,During
R5019	T16825	T16823	punct	", ",abolished
R502	T3387	T3388	det	the,patterns
R5020	T16826	T16827	det	the,site
R5021	T16827	T16823	nsubjpass	site,abolished
R5022	T16828	T16827	compound	BamHI,site
R5023	T16829	T16827	prep	at,site
R5024	T16830	T16831	det	the,end
R5025	T16831	T16829	pobj	end,at
R5026	T16832	T16831	nummod	3,end
R5027	T16833	T16832	punct	',3
R5028	T16834	T16831	prep	of,end
R5029	T16835	T16836	det	the,arm
R503	T3513	T3514	det	these,enzymes
R5030	T16836	T16834	pobj	arm,of
R5031	T16837	T16836	amod	short,arm
R5032	T16838	T16823	auxpass	was,abolished
R5033	T16839	T16823	punct	.,abolished
R5034	T16841	T16842	det	The,loss
R5035	T16842	T16843	nsubj	loss,provided
R5036	T16844	T16842	prep	of,loss
R5037	T16845	T16846	det	this,site
R5038	T16846	T16844	pobj	site,of
R5039	T16847	T16846	compound	BamHI,site
R504	T3514	T3512	pobj	enzymes,of
R5040	T16848	T16842	prep	in,loss
R5041	T16849	T16850	det	the,allele
R5042	T16850	T16848	pobj	allele,in
R5043	T16851	T16850	compound	mutant,allele
R5044	T16852	T16842	punct	", ",loss
R5045	T16853	T16842	acl	coupled,loss
R5046	T16854	T16853	prep	with,coupled
R5047	T16855	T16856	det	the,introduction
R5048	T16856	T16854	pobj	introduction,with
R5049	T16857	T16856	prep	of,introduction
R505	T3515	T3516	punct	[,3
R5050	T16858	T16859	det	a,site
R5051	T16859	T16857	pobj	site,of
R5052	T16860	T16859	amod	new,site
R5053	T16861	T16859	compound	BamHI,site
R5054	T16862	T16853	prep	at,coupled
R5055	T16863	T16864	det	the,end
R5056	T16864	T16862	pobj	end,at
R5057	T16865	T16864	nummod	3,end
R5058	T16866	T16865	punct	',3
R5059	T16867	T16864	prep	of,end
R506	T3388	T3383	nsubj	patterns,suggest
R5060	T16868	T16869	det	the,cassette
R5061	T16869	T16867	pobj	cassette,of
R5062	T16870	T16871	compound	PGK,Neo
R5063	T16871	T16869	compound	Neo,cassette
R5064	T16872	T16871	punct	-,Neo
R5065	T16873	T16843	punct	", ",provided
R5066	T16874	T16875	det	a,basis
R5067	T16875	T16843	dobj	basis,provided
R5068	T16876	T16875	prep	for,basis
R5069	T16877	T16876	pcomp	distinguishing,for
R507	T3516	T3504	parataxis	3,contribute
R5070	T16878	T16879	det	the,alleles
R5071	T16879	T16877	dobj	alleles,distinguishing
R5072	T16880	T16881	amod	wild,type
R5073	T16881	T16879	nmod	type,alleles
R5074	T16882	T16881	punct	-,type
R5075	T16883	T16881	cc	and,type
R5076	T16884	T16881	conj	mutant,type
R5077	T16885	T16877	prep	by,distinguishing
R5078	T16886	T16887	compound	Southern,blotting
R5079	T16887	T16885	pobj	blotting,by
R508	T3517	T3516	punct	],3
R5080	T16888	T16843	punct	.,provided
R5081	T16890	T16891	det	The,arm
R5082	T16891	T16896	nsubjpass	arm,provided
R5083	T16892	T16891	amod	long,arm
R5084	T16893	T16891	punct	(,arm
R5085	T16894	T16891	amod	downstream,arm
R5086	T16895	T16891	punct	),arm
R5087	T16897	T16891	prep	of,arm
R5088	T16898	T16897	pobj	homology,of
R5089	T16899	T16896	auxpass	was,provided
R509	T3518	T3484	punct	.,assumed
R5090	T16900	T16896	agent	by,provided
R5091	T16901	T16902	det	a,fragment
R5092	T16902	T16900	pobj	fragment,by
R5093	T16903	T16904	nummod	7,kb
R5094	T16904	T16902	compound	kb,fragment
R5095	T16905	T16904	punct	-,kb
R5096	T16906	T16907	compound	HindIII,KpnI
R5097	T16907	T16902	compound	KpnI,fragment
R5098	T16908	T16907	punct	-,KpnI
R5099	T16909	T16902	punct	", ",fragment
R51	T723	T720	appos	calpain,Capn1
R510	T3520	T3521	prep	Without,presents
R5100	T16910	T16902	acl	extending,fragment
R5101	T16911	T16910	prep	from,extending
R5102	T16912	T16911	pobj	intron,from
R5103	T16913	T16912	nummod	7,intron
R5104	T16914	T16911	prep	to,from
R5105	T16915	T16914	pobj	intron,to
R5106	T16916	T16915	nummod	12,intron
R5107	T16917	T16902	punct	", ",fragment
R5108	T16918	T16902	acl	inserted,fragment
R5109	T16919	T16918	prep	between,inserted
R511	T3389	T3388	amod	differential,patterns
R5110	T16920	T16921	det	the,cassettes
R5111	T16921	T16919	pobj	cassettes,between
R5112	T16922	T16923	nmod	PGK,Neo
R5113	T16923	T16921	nmod	Neo,cassettes
R5114	T16924	T16923	punct	-,Neo
R5115	T16925	T16923	cc	and,Neo
R5116	T16926	T16927	compound	thymidine,kinase
R5117	T16927	T16923	conj	kinase,Neo
R5118	T16928	T16927	punct	(,kinase
R5119	T16929	T16927	appos	tk,kinase
R512	T3522	T3523	nmod	apriori,knowledge
R5120	T16930	T16921	punct	),cassettes
R5121	T16931	T16896	punct	.,provided
R5122	T17462	T17463	compound	ES,culture
R5123	T17464	T17463	compound	cell,culture
R5124	T17465	T17463	punct	", ",culture
R5125	T17466	T17463	conj	transfection,culture
R5126	T17467	T17466	cc	and,transfection
R5127	T17468	T17466	conj	selection,transfection
R5128	T17469	T17468	prep	of,selection
R5129	T17470	T17471	amod	targeted,clones
R513	T3523	T3520	pobj	knowledge,Without
R5130	T17471	T17469	pobj	clones,of
R5131	T17473	T17474	compound	Mouse,cells
R5132	T17474	T17477	nsubjpass	cells,maintained
R5133	T17475	T17474	compound	R1,cells
R5134	T17476	T17474	compound	ES,cells
R5135	T17478	T17479	punct	[,49
R5136	T17479	T17474	parataxis	49,cells
R5137	T17480	T17479	punct	],49
R5138	T17481	T17477	auxpass	were,maintained
R5139	T17482	T17477	prep	on,maintained
R514	T3390	T3388	compound	expression,patterns
R5140	T17483	T17484	npadvmod	gelatin,coated
R5141	T17484	T17486	amod	coated,plates
R5142	T17485	T17484	punct	-,coated
R5143	T17486	T17482	pobj	plates,on
R5144	T17487	T17477	prep	with,maintained
R5145	T17488	T17489	compound	feeder,layers
R5146	T17489	T17487	pobj	layers,with
R5147	T17490	T17489	prep	of,layers
R5148	T17491	T17492	nmod	mouse,fibroblasts
R5149	T17492	T17490	pobj	fibroblasts,of
R515	T3524	T3523	prep	of,knowledge
R5150	T17493	T17492	amod	embryonic,fibroblasts
R5151	T17494	T17477	prep	at,maintained
R5152	T17495	T17496	nummod	37,°C
R5153	T17496	T17494	pobj	°C,at
R5154	T17497	T17477	prep	under,maintained
R5155	T17498	T17499	nummod	5,%
R5156	T17499	T17500	compound	%,CO2
R5157	T17500	T17497	pobj	CO2,under
R5158	T17501	T17477	prep	in,maintained
R5159	T17502	T17503	compound	ES,medium
R516	T3525	T3526	det	the,factors
R5160	T17503	T17501	pobj	medium,in
R5161	T17504	T17503	compound	cell,medium
R5162	T17505	T17506	punct	(,DMEM
R5163	T17506	T17477	parataxis	DMEM,maintained
R5164	T17507	T17508	punct	[,glucose
R5165	T17508	T17506	parataxis	glucose,DMEM
R5166	T17509	T17508	amod	high,glucose
R5167	T17510	T17506	punct	],DMEM
R5168	T17511	T17506	acl	supplemented,DMEM
R5169	T17512	T17511	prep	with,supplemented
R517	T3526	T3524	pobj	factors,of
R5170	T17513	T17514	nummod	15,%
R5171	T17514	T17515	nmod	%,serum
R5172	T17515	T17512	pobj	serum,with
R5173	T17516	T17515	amod	fetal,serum
R5174	T17517	T17515	amod	bovine,serum
R5175	T17518	T17515	punct	", ",serum
R5176	T17519	T17520	nummod	0.1,mM
R5177	T17520	T17521	nmod	mM,acids
R5178	T17521	T17515	conj	acids,serum
R5179	T17522	T17521	amod	non-essential,acids
R518	T3391	T3388	prep	of,patterns
R5180	T17523	T17521	compound	amino,acids
R5181	T17524	T17521	punct	", ",acids
R5182	T17525	T17526	nummod	2,mM
R5183	T17526	T17527	compound	mM,glutamine
R5184	T17527	T17521	conj	glutamine,acids
R5185	T17528	T17527	punct	", ",glutamine
R5186	T17529	T17530	nummod	1,mM
R5187	T17530	T17531	compound	mM,pyruvate
R5188	T17531	T17527	conj	pyruvate,glutamine
R5189	T17532	T17531	compound	sodium,pyruvate
R519	T3527	T3526	acl	regulating,factors
R5190	T17533	T17531	punct	", ",pyruvate
R5191	T17534	T17535	nummod	0.1,mM
R5192	T17535	T17536	nmod	mM,mercaptoethanol
R5193	T17536	T17531	conj	mercaptoethanol,pyruvate
R5194	T17537	T17536	nummod	2,mercaptoethanol
R5195	T17538	T17536	punct	-,mercaptoethanol
R5196	T17539	T17536	punct	", ",mercaptoethanol
R5197	T17540	T17541	nummod	100,U
R5198	T17541	T17542	nmod	U,penicillin
R5199	T17542	T17536	conj	penicillin,mercaptoethanol
R52	T724	T723	punct	-,calpain
R520	T3528	T3529	compound	calpain,activity
R5200	T17543	T17544	punct	/,ml
R5201	T17544	T17541	prep	ml,U
R5202	T17545	T17542	punct	", ",penicillin
R5203	T17546	T17547	nummod	100,μg
R5204	T17547	T17548	nmod	μg,streptomycin
R5205	T17548	T17542	conj	streptomycin,penicillin
R5206	T17549	T17550	punct	/,ml
R5207	T17550	T17547	prep	ml,μg
R5208	T17551	T17548	punct	", ",streptomycin
R5209	T17552	T17553	nummod	25,μg
R521	T3529	T3527	dobj	activity,regulating
R5210	T17553	T17554	nmod	μg,amphotericin
R5211	T17554	T17548	conj	amphotericin,streptomycin
R5212	T17555	T17556	punct	/,ml
R5213	T17556	T17553	prep	ml,μg
R5214	T17557	T17554	punct	", ",amphotericin
R5215	T17558	T17554	cc	and,amphotericin
R5216	T17559	T17560	nummod	"1,000",U
R5217	T17560	T17554	conj	U,amphotericin
R5218	T17561	T17562	punct	/,ml
R5219	T17562	T17560	prep	ml,U
R522	T3392	T3393	nmod	μ,calpain
R5220	T17563	T17560	prep	of,U
R5221	T17564	T17563	pobj	ESGro,of
R5222	T17565	T17566	punct	[,Chemicon
R5223	T17566	T17564	parataxis	Chemicon,ESGro
R5224	T17567	T17566	punct	],Chemicon
R5225	T17568	T17477	punct	),maintained
R5226	T17569	T17477	punct	.,maintained
R5227	T17571	T17572	amod	Fetal,serum
R5228	T17572	T17574	nsubjpass	serum,tested
R5229	T17573	T17572	amod	bovine,serum
R523	T3530	T3526	cc	or,factors
R5230	T17575	T17572	prep	from,serum
R5231	T17576	T17577	compound	HyClone,Inc
R5232	T17577	T17575	pobj	Inc,from
R5233	T17578	T17577	compound	Laboratories,Inc
R5234	T17579	T17580	punct	(,Logan
R5235	T17580	T17577	parataxis	Logan,Inc
R5236	T17581	T17580	punct	", ",Logan
R5237	T17582	T17580	npadvmod	Utah,Logan
R5238	T17583	T17580	punct	),Logan
R5239	T17584	T17574	auxpass	was,tested
R524	T3531	T3532	poss	their,substrates
R5240	T17585	T17574	prep	for,tested
R5241	T17586	T17587	poss	its,ability
R5242	T17587	T17585	pobj	ability,for
R5243	T17588	T17589	aux	to,support
R5244	T17589	T17587	acl	support,ability
R5245	T17590	T17591	compound	ES,cell
R5246	T17591	T17592	compound	cell,growth
R5247	T17592	T17589	dobj	growth,support
R5248	T17593	T17574	punct	.,tested
R5249	T17595	T17596	nsubj	Gelatin,was
R525	T3532	T3526	conj	substrates,factors
R5250	T17597	T17596	prep	from,was
R5251	T17598	T17599	compound	Sigma,Aldrich
R5252	T17599	T17601	compound	Aldrich,Canada
R5253	T17600	T17599	punct	-,Aldrich
R5254	T17601	T17597	pobj	Canada,from
R5255	T17602	T17603	punct	(,Oakville
R5256	T17603	T17596	parataxis	Oakville,was
R5257	T17604	T17603	punct	", ",Oakville
R5258	T17605	T17603	npadvmod	Ontario,Oakville
R5259	T17606	T17603	punct	),Oakville
R526	T3393	T3391	pobj	calpain,of
R5260	T17607	T17596	punct	.,was
R5261	T17609	T17610	mark	Unless,specified
R5262	T17610	T17612	advcl	specified,were
R5263	T17611	T17610	advmod	otherwise,specified
R5264	T17613	T17612	punct	", ",were
R5265	T17614	T17615	det	all,reagents
R5266	T17615	T17612	nsubj	reagents,were
R5267	T17616	T17615	amod	other,reagents
R5268	T17617	T17618	compound	tissue,culture
R5269	T17618	T17615	compound	culture,reagents
R527	T3533	T3532	amod	relevant,substrates
R5270	T17619	T17612	prep	from,were
R5271	T17620	T17621	compound	Gibco,BRL
R5272	T17621	T17619	pobj	BRL,from
R5273	T17622	T17621	punct	-,BRL
R5274	T17623	T17612	punct	.,were
R5275	T17625	T17626	det	The,construct
R5276	T17626	T17628	nsubjpass	construct,linearized
R5277	T17627	T17626	compound	targeting,construct
R5278	T17629	T17628	auxpass	was,linearized
R5279	T17630	T17628	agent	by,linearized
R528	T3534	T3521	punct	", ",presents
R5280	T17631	T17632	compound	NotI,digestion
R5281	T17632	T17630	pobj	digestion,by
R5282	T17633	T17628	cc	and,linearized
R5283	T17634	T17628	conj	electroporated,linearized
R5284	T17635	T17634	prep	into,electroporated
R5285	T17636	T17637	compound	R1,cells
R5286	T17637	T17635	pobj	cells,into
R5287	T17638	T17637	compound	ES,cells
R5288	T17639	T17628	punct	.,linearized
R5289	T17641	T17642	nsubjpass	Cells,plated
R529	T3394	T3392	punct	-,μ
R5290	T17643	T17642	auxpass	were,plated
R5291	T17644	T17642	prep	without,plated
R5292	T17645	T17646	compound	feeder,layers
R5293	T17646	T17644	pobj	layers,without
R5294	T17647	T17642	prep	on,plated
R5295	T17648	T17649	npadvmod	gelatin,coated
R5296	T17649	T17651	amod	coated,plates
R5297	T17650	T17649	punct	-,coated
R5298	T17651	T17647	pobj	plates,on
R5299	T17652	T17642	cc	and,plated
R53	T725	T720	punct	),Capn1
R530	T3535	T3521	nsubj	elucidation,presents
R5300	T17653	T17654	amod	transformed,clones
R5301	T17654	T17655	nsubjpass	clones,selected
R5302	T17655	T17642	conj	selected,plated
R5303	T17656	T17655	auxpass	were,selected
R5304	T17657	T17655	prep	in,selected
R5305	T17658	T17659	det	the,presence
R5306	T17659	T17657	pobj	presence,in
R5307	T17660	T17659	prep	of,presence
R5308	T17661	T17662	nummod	200,μg
R5309	T17662	T17663	nmod	μg,G418
R531	T3395	T3392	cc	and,μ
R5310	T17663	T17660	pobj	G418,of
R5311	T17664	T17665	punct	/,ml
R5312	T17665	T17662	prep	ml,μg
R5313	T17666	T17667	punct	(,BRL
R5314	T17667	T17663	parataxis	BRL,G418
R5315	T17668	T17667	compound	Gibco,BRL
R5316	T17669	T17667	punct	-,BRL
R5317	T17670	T17667	punct	),BRL
R5318	T17671	T17663	cc	and,G418
R5319	T17672	T17673	nummod	2,μM
R532	T3536	T3535	prep	of,elucidation
R5320	T17673	T17674	compound	μM,ganciclovir
R5321	T17674	T17663	conj	ganciclovir,G418
R5322	T17675	T17676	punct	(,Syntex
R5323	T17676	T17674	parataxis	Syntex,ganciclovir
R5324	T17677	T17676	punct	", ",Syntex
R5325	T17678	T17676	amod	Inc.,Syntex
R5326	T17679	T17676	punct	),Syntex
R5327	T17680	T17655	prep	for,selected
R5328	T17681	T17682	nummod	eight,days
R5329	T17682	T17680	pobj	days,for
R533	T3537	T3538	amod	biological,functions
R5330	T17683	T17655	punct	.,selected
R5331	T17685	T17686	npadvmod	Drug,resistant
R5332	T17686	T17688	amod	resistant,clones
R5333	T17687	T17686	punct	-,resistant
R5334	T17688	T17689	nsubjpass	clones,picked
R5335	T17690	T17689	auxpass	were,picked
R5336	T17691	T17689	punct	", ",picked
R5337	T17692	T17689	conj	expanded,picked
R5338	T17693	T17692	prep	on,expanded
R5339	T17694	T17695	npadvmod	gelatin,coated
R534	T3538	T3536	pobj	functions,of
R5340	T17695	T17697	amod	coated,plates
R5341	T17696	T17695	punct	-,coated
R5342	T17697	T17693	pobj	plates,on
R5343	T17698	T17692	punct	", ",expanded
R5344	T17699	T17692	cc	and,expanded
R5345	T17700	T17692	conj	genotyped,expanded
R5346	T17701	T17700	prep	by,genotyped
R5347	T17702	T17703	compound	Southern,blotting
R5348	T17703	T17701	pobj	blotting,by
R5349	T17704	T17703	cc	and,blotting
R535	T3539	T3535	prep	for,elucidation
R5350	T17705	T17706	compound	PCR,analysis
R5351	T17706	T17703	conj	analysis,blotting
R5352	T17707	T17708	punct	(,see
R5353	T17708	T17700	parataxis	see,genotyped
R5354	T17709	T17708	advmod	below,see
R5355	T17710	T17708	punct	),see
R5356	T17711	T17689	punct	.,picked
R5357	T17989	T17988	prep	of,Generation
R5358	T17990	T17991	amod	targeted,mice
R5359	T17991	T17989	pobj	mice,of
R536	T3396	T3392	conj	m,μ
R5360	T17993	T17994	nmod	Capn2,cells
R5361	T17994	T17999	nsubjpass	cells,aggregated
R5362	T17995	T17993	punct	+,Capn2
R5363	T17996	T17993	punct	/,Capn2
R5364	T17997	T17993	punct	-,Capn2
R5365	T17998	T17994	compound	ES,cells
R5366	T18000	T17999	auxpass	were,aggregated
R5367	T18001	T17999	advmod	overnight,aggregated
R5368	T18002	T17999	prep	with,aggregated
R5369	T18003	T18004	nummod	8,cell
R537	T3540	T3539	pobj	calpains,for
R5370	T18004	T18006	compound	cell,embryos
R5371	T18005	T18004	punct	-,cell
R5372	T18006	T18002	pobj	embryos,with
R5373	T18007	T18006	acl	recovered,embryos
R5374	T18008	T18007	prep	from,recovered
R5375	T18009	T18010	compound	CD1,matings
R5376	T18010	T18008	pobj	matings,from
R5377	T18011	T17999	punct	", ",aggregated
R5378	T18012	T18013	mark	as,described
R5379	T18013	T17999	advcl	described,aggregated
R538	T3541	T3542	det	a,challenge
R5380	T18014	T18013	advmod	previously,described
R5381	T18015	T18016	punct	[,31
R5382	T18016	T17999	parataxis	31,aggregated
R5383	T18017	T18016	punct	],31
R5384	T18018	T17999	punct	.,aggregated
R5385	T18020	T18021	prep	On,transferred
R5386	T18022	T18023	det	the,day
R5387	T18023	T18020	pobj	day,On
R5388	T18024	T18023	amod	next,day
R5389	T18025	T18021	punct	", ",transferred
R539	T3542	T3521	dobj	challenge,presents
R5390	T18026	T18021	nsubjpass	blastocysts,transferred
R5391	T18027	T18021	auxpass	were,transferred
R5392	T18028	T18021	prep	to,transferred
R5393	T18029	T18030	amod	pseudopregnant,females
R5394	T18030	T18028	pobj	females,to
R5395	T18031	T18030	compound	CD1,females
R5396	T18032	T18021	punct	.,transferred
R5397	T18034	T18035	amod	Chimeric,animals
R5398	T18035	T18036	nsubj	animals,were
R5399	T18037	T18036	acomp	identifiable,were
R54	T726	T720	cc	or,Capn1
R540	T3397	T3393	punct	-,calpain
R5400	T18038	T18036	prep	at,were
R5401	T18039	T18038	pobj	birth,at
R5402	T18040	T18036	prep	by,were
R5403	T18041	T18042	amod	black,pigmentation
R5404	T18042	T18040	pobj	pigmentation,by
R5405	T18043	T18042	compound	eye,pigmentation
R5406	T18044	T18040	cc	and,by
R5407	T18045	T18046	advmod	subsequently,by
R5408	T18046	T18040	conj	by,by
R5409	T18047	T18046	pobj	patches,by
R541	T3543	T3521	punct	.,presents
R5410	T18048	T18047	prep	of,patches
R5411	T18049	T18050	compound	agouti,colour
R5412	T18050	T18048	pobj	colour,of
R5413	T18051	T18050	compound	coat,colour
R5414	T18052	T18036	punct	.,were
R5415	T18054	T18055	amod	Chimeric,males
R5416	T18055	T18056	nsubjpass	males,bred
R5417	T18057	T18056	auxpass	were,bred
R5418	T18058	T18056	prep	with,bred
R5419	T18059	T18060	compound	CD1,females
R542	T3398	T3388	prep	in,patterns
R5420	T18060	T18058	pobj	females,with
R5421	T18061	T18062	aux	to,identify
R5422	T18062	T18056	advcl	identify,bred
R5423	T18063	T18064	det	those,males
R5424	T18064	T18062	dobj	males,identify
R5425	T18065	T18064	amod	capable,males
R5426	T18066	T18065	prep	of,capable
R5427	T18067	T18068	compound	germline,transmission
R5428	T18068	T18066	pobj	transmission,of
R5429	T18069	T18056	punct	.,bred
R543	T3545	T3546	nsubj	Research,linked
R5430	T18071	T18072	nsubjpass	These,bred
R5431	T18073	T18072	auxpass	were,bred
R5432	T18074	T18072	advmod	then,bred
R5433	T18075	T18072	prep	with,bred
R5434	T18076	T18077	compound	129SvJ,females
R5435	T18077	T18075	pobj	females,with
R5436	T18078	T18079	aux	to,establish
R5437	T18079	T18072	advcl	establish,bred
R5438	T18080	T18081	det	the,mutation
R5439	T18081	T18079	dobj	mutation,establish
R544	T3547	T3545	prep	on,Research
R5440	T18082	T18079	prep	in,establish
R5441	T18083	T18084	det	an,background
R5442	T18084	T18082	pobj	background,in
R5443	T18085	T18084	amod	inbred,background
R5444	T18086	T18084	amod	genetic,background
R5445	T18087	T18072	punct	.,bred
R5446	T18089	T18090	compound	Mouse,protocols
R5447	T18090	T18091	nsubjpass	protocols,approved
R5448	T18092	T18091	auxpass	were,approved
R5449	T18093	T18091	agent	by,approved
R545	T3399	T3400	amod	mammalian,tissues
R5450	T18094	T18095	det	the,Queen
R5451	T18095	T18096	poss	Queen,University
R5452	T18096	T18098	compound	University,Committee
R5453	T18097	T18095	case	's,Queen
R5454	T18098	T18093	pobj	Committee,by
R5455	T18099	T18100	compound	Animal,Care
R5456	T18100	T18098	compound	Care,Committee
R5457	T18101	T18091	prep	according,approved
R5458	T18102	T18101	prep	to,according
R5459	T18103	T18104	det	the,guidelines
R546	T3548	T3547	pobj	calpains,on
R5460	T18104	T18102	pobj	guidelines,to
R5461	T18105	T18104	prep	of,guidelines
R5462	T18106	T18107	det	the,Council
R5463	T18107	T18105	pobj	Council,of
R5464	T18108	T18107	compound	Canadian,Council
R5465	T18109	T18107	prep	on,Council
R5466	T18110	T18111	compound	Animal,Care
R5467	T18111	T18109	pobj	Care,on
R5468	T18112	T18091	punct	.,approved
R5469	T19530	T19531	compound	Genotyping,methods
R547	T3549	T3546	aux	has,linked
R5470	T19533	T19534	amod	Several,blot
R5471	T19534	T19536	nsubjpass	blot,exploited
R5472	T19535	T19534	compound	Southern,blot
R5473	T19537	T19534	cc	and,blot
R5474	T19538	T19539	compound	PCR,strategies
R5475	T19539	T19534	conj	strategies,blot
R5476	T19540	T19536	auxpass	were,exploited
R5477	T19541	T19536	prep	in,exploited
R5478	T19542	T19541	pobj	order,in
R5479	T19543	T19544	aux	to,determine
R548	T3550	T3546	dobj	them,linked
R5480	T19544	T19542	acl	determine,order
R5481	T19545	T19546	det	the,genotype
R5482	T19546	T19544	dobj	genotype,determine
R5483	T19547	T19546	prep	of,genotype
R5484	T19548	T19549	det	the,locus
R5485	T19549	T19547	pobj	locus,of
R5486	T19550	T19549	compound	Capn2,locus
R5487	T19551	T19536	punct	.,exploited
R5488	T19553	T19554	compound	Southern,blotting
R5489	T19554	T19555	nsubjpass	blotting,carried
R549	T3400	T3398	pobj	tissues,in
R5490	T19556	T19555	auxpass	was,carried
R5491	T19557	T19555	prt	out,carried
R5492	T19558	T19555	advcl	using,carried
R5493	T19559	T19560	det	the,system
R5494	T19560	T19558	dobj	system,using
R5495	T19561	T19560	nmod	digoxigenin,system
R5496	T19562	T19561	punct	(,digoxigenin
R5497	T19563	T19561	appos	DIG,digoxigenin
R5498	T19564	T19560	punct	),system
R5499	T19565	T19560	amod	non-radioactive,system
R55	T727	T720	conj	Capn2,Capn1
R550	T3551	T3546	prep	with,linked
R5500	T19566	T19567	punct	(,Roche
R5501	T19567	T19560	parataxis	Roche,system
R5502	T19568	T19567	punct	),Roche
R5503	T19569	T19555	punct	.,carried
R5504	T19571	T19572	prep	In,blotted
R5505	T19573	T19574	amod	most,cases
R5506	T19574	T19571	pobj	cases,In
R5507	T19575	T19572	punct	", ",blotted
R5508	T19576	T19572	nsubjpass	membranes,blotted
R5509	T19577	T19572	auxpass	were,blotted
R551	T3552	T3553	det	a,variety
R5510	T19578	T19572	prep	with,blotted
R5511	T19579	T19580	npadvmod	BamHI,digested
R5512	T19580	T19582	amod	digested,DNA
R5513	T19581	T19580	punct	-,digested
R5514	T19582	T19578	pobj	DNA,with
R5515	T19583	T19582	amod	genomic,DNA
R5516	T19584	T19572	cc	and,blotted
R5517	T19585	T19572	conj	hybridized,blotted
R5518	T19586	T19585	prep	with,hybridized
R5519	T19587	T19588	det	a,fragment
R552	T3401	T3402	nsubj	they,have
R5520	T19588	T19586	pobj	fragment,with
R5521	T19589	T19590	npadvmod	DIG,labeled
R5522	T19590	T19588	amod	labeled,fragment
R5523	T19591	T19590	punct	-,labeled
R5524	T19592	T19593	nummod	823,bp
R5525	T19593	T19588	nmod	bp,fragment
R5526	T19594	T19595	npadvmod	exon,containing
R5527	T19595	T19588	amod	containing,fragment
R5528	T19596	T19594	nummod	4,exon
R5529	T19597	T19595	punct	-,containing
R553	T3553	T3551	pobj	variety,with
R5530	T19598	T19599	compound	BamHI,HindIII
R5531	T19599	T19588	compound	HindIII,fragment
R5532	T19600	T19599	punct	-,HindIII
R5533	T19601	T19588	acl	located,fragment
R5534	T19602	T19603	advmod	immediately,upstream
R5535	T19603	T19601	advmod	upstream,located
R5536	T19604	T19603	prep	of,upstream
R5537	T19605	T19606	det	the,arm
R5538	T19606	T19604	pobj	arm,of
R5539	T19607	T19606	amod	short,arm
R554	T3554	T3553	amod	wide,variety
R5540	T19608	T19606	prep	of,arm
R5541	T19609	T19608	pobj	homology,of
R5542	T19610	T19611	punct	(,Figure
R5543	T19611	T19585	parataxis	Figure,hybridized
R5544	T19612	T19611	nummod	1,Figure
R5545	T19613	T19611	punct	),Figure
R5546	T19614	T19572	punct	.,blotted
R5547	T19616	T19617	det	A,fragment
R5548	T19617	T19623	nsubjpass	fragment,used
R5549	T19618	T19619	nummod	681,bp
R555	T3555	T3553	prep	of,variety
R5550	T19619	T19617	compound	bp,fragment
R5551	T19620	T19621	compound	PstI,XbaI
R5552	T19621	T19617	compound	XbaI,fragment
R5553	T19622	T19621	punct	-,XbaI
R5554	T19624	T19617	prep	from,fragment
R5555	T19625	T19626	det	the,cassette
R5556	T19626	T19624	pobj	cassette,from
R5557	T19627	T19628	compound	PGK,Neo
R5558	T19628	T19626	compound	Neo,cassette
R5559	T19629	T19628	punct	-,Neo
R556	T3402	T3383	advcl	have,suggest
R5560	T19630	T19623	auxpass	was,used
R5561	T19631	T19623	advmod	also,used
R5562	T19632	T19633	aux	to,probe
R5563	T19633	T19623	advcl	probe,used
R5564	T19634	T19635	compound	Southern,blots
R5565	T19635	T19633	dobj	blots,probe
R5566	T19636	T19623	prep	in,used
R5567	T19637	T19636	pobj	order,in
R5568	T19638	T19639	aux	to,verify
R5569	T19639	T19637	acl	verify,order
R557	T3556	T3555	pobj	functions,of
R5570	T19640	T19641	det	a,event
R5571	T19641	T19639	dobj	event,verify
R5572	T19642	T19641	amod	single,event
R5573	T19643	T19641	compound	integration,event
R5574	T19644	T19623	prep	in,used
R5575	T19645	T19646	amod	targeted,clones
R5576	T19646	T19644	pobj	clones,in
R5577	T19647	T19623	punct	.,used
R5578	T19649	T19650	nsubjpass	Genotyping,carried
R5579	T19651	T19650	auxpass	was,carried
R558	T3557	T3553	prep	including,variety
R5580	T19652	T19650	advmod	also,carried
R5581	T19653	T19650	prt	out,carried
R5582	T19654	T19650	agent	by,carried
R5583	T19655	T19656	compound	PCR,analysis
R5584	T19656	T19654	pobj	analysis,by
R5585	T19657	T19656	prep	of,analysis
R5586	T19658	T19659	amod	genomic,DNA
R5587	T19659	T19657	pobj	DNA,of
R5588	T19660	T19650	punct	.,carried
R5589	T19662	T19663	det	The,sequences
R559	T3558	T3559	compound	muscle,growth
R5590	T19663	T19664	nsubjpass	sequences,listed
R5591	T19665	T19663	prep	of,sequences
R5592	T19666	T19667	det	all,primers
R5593	T19667	T19665	pobj	primers,of
R5594	T19668	T19667	compound	oligonucleotide,primers
R5595	T19669	T19664	auxpass	are,listed
R5596	T19670	T19664	prep	in,listed
R5597	T19671	T19670	pobj	Table,in
R5598	T19672	T19671	nummod	1,Table
R5599	T19673	T19664	punct	.,listed
R56	T728	T727	punct	(,Capn2
R560	T3403	T3404	det	some,functions
R5600	T19675	T19676	det	A,strategy
R5601	T19676	T19681	nsubj	strategy,was
R5602	T19677	T19678	amod	single,step
R5603	T19678	T19676	compound	step,strategy
R5604	T19679	T19678	punct	-,step
R5605	T19680	T19676	compound	PCR,strategy
R5606	T19682	T19681	acomp	sufficient,was
R5607	T19683	T19682	prep	for,sufficient
R5608	T19684	T19683	pcomp	genotyping,for
R5609	T19685	T19686	compound	ES,cells
R561	T3559	T3557	pobj	growth,including
R5610	T19686	T19684	dobj	cells,genotyping
R5611	T19687	T19686	cc	or,cells
R5612	T19688	T19686	conj	biopsies,cells
R5613	T19689	T19688	prep	from,biopsies
R5614	T19690	T19691	amod	post-implantation,embryos
R5615	T19691	T19689	pobj	embryos,from
R5616	T19692	T19691	cc	and,embryos
R5617	T19693	T19691	conj	weanlings,embryos
R5618	T19694	T19695	punct	(,Figure
R5619	T19695	T19684	parataxis	Figure,genotyping
R562	T3404	T3402	dobj	functions,have
R5620	T19696	T19695	nummod	1,Figure
R5621	T19697	T19695	punct	),Figure
R5622	T19698	T19681	punct	.,was
R5623	T19700	T19701	det	A,segment
R5624	T19701	T19704	nsubjpass	segment,amplified
R5625	T19702	T19703	nummod	"2,748",bp
R5626	T19703	T19701	compound	bp,segment
R5627	T19705	T19701	prep	of,segment
R5628	T19706	T19707	det	the,allele
R5629	T19707	T19705	pobj	allele,of
R563	T3560	T3559	punct	", ",growth
R5630	T19708	T19709	amod	wild,type
R5631	T19709	T19707	compound	type,allele
R5632	T19710	T19709	punct	-,type
R5633	T19711	T19701	cc	and,segment
R5634	T19712	T19713	det	a,segment
R5635	T19713	T19701	conj	segment,segment
R5636	T19714	T19715	nummod	"2,711",bp
R5637	T19715	T19713	compound	bp,segment
R5638	T19716	T19713	prep	of,segment
R5639	T19717	T19718	det	the,allele
R564	T3405	T3406	npadvmod	isoform,specific
R5640	T19718	T19716	pobj	allele,of
R5641	T19719	T19718	compound	mutant,allele
R5642	T19720	T19704	auxpass	were,amplified
R5643	T19721	T19704	prep	in,amplified
R5644	T19722	T19723	amod	separate,reactions
R5645	T19723	T19721	pobj	reactions,in
R5646	T19724	T19704	advcl	using,amplified
R5647	T19725	T19726	det	a,primer
R5648	T19726	T19724	dobj	primer,using
R5649	T19727	T19728	amod	common,sense
R565	T3561	T3559	conj	development,growth
R5650	T19728	T19726	compound	sense,primer
R5651	T19729	T19730	punct	(,intron
R5652	T19730	T19728	parataxis	intron,sense
R5653	T19731	T19730	nummod	4,intron
R5654	T19732	T19730	punct	),intron
R5655	T19733	T19726	punct	", ",primer
R5656	T19734	T19726	acl	located,primer
R5657	T19735	T19734	prep	outside,located
R5658	T19736	T19737	det	the,arm
R5659	T19737	T19735	pobj	arm,outside
R566	T3562	T3561	punct	", ",development
R5660	T19738	T19737	amod	short,arm
R5661	T19739	T19737	prep	of,arm
R5662	T19740	T19739	pobj	homology,of
R5663	T19741	T19726	punct	", ",primer
R5664	T19742	T19726	cc	and,primer
R5665	T19743	T19744	amod	distinct,primers
R5666	T19744	T19726	conj	primers,primer
R5667	T19745	T19744	compound	antisense,primers
R5668	T19746	T19747	dep	which,hybridized
R5669	T19747	T19744	relcl	hybridized,primers
R567	T3563	T3561	conj	degeneration,development
R5670	T19748	T19747	prep	to,hybridized
R5671	T19749	T19750	preconj	either,type
R5672	T19750	T19753	nmod	type,sequence
R5673	T19751	T19750	amod	wild,type
R5674	T19752	T19750	punct	-,type
R5675	T19753	T19748	pobj	sequence,to
R5676	T19754	T19755	punct	(,exon
R5677	T19755	T19750	parataxis	exon,type
R5678	T19756	T19755	nummod	7,exon
R5679	T19757	T19755	punct	),exon
R568	T3406	T3404	amod	specific,functions
R5680	T19758	T19750	cc	or,type
R5681	T19759	T19750	conj	mutant,type
R5682	T19760	T19761	punct	(,Neo
R5683	T19761	T19759	parataxis	Neo,mutant
R5684	T19762	T19761	compound	PGK,Neo
R5685	T19763	T19761	punct	-,Neo
R5686	T19764	T19761	punct	),Neo
R5687	T19765	T19766	punct	(,I
R5688	T19766	T19747	parataxis	I,hybridized
R5689	T19767	T19766	compound	Table,I
R569	T3564	T3565	punct	(,3
R5690	T19768	T19766	punct	),I
R5691	T19769	T19704	punct	.,amplified
R5692	T19771	T19772	det	The,parameters
R5693	T19772	T19774	nsubj	parameters,included
R5694	T19773	T19772	amod	thermocycling,parameters
R5695	T19775	T19776	det	a,step
R5696	T19776	T19774	dobj	step,included
R5697	T19777	T19778	nummod	five,minute
R5698	T19778	T19776	nmod	minute,step
R5699	T19779	T19776	amod	initial,step
R57	T729	T730	compound	m,calpain
R570	T3407	T3406	cc	and,specific
R5700	T19780	T19776	compound	denaturation,step
R5701	T19781	T19776	prep	at,step
R5702	T19782	T19783	nummod	95,°C
R5703	T19783	T19781	pobj	°C,at
R5704	T19784	T19776	punct	", ",step
R5705	T19785	T19786	nummod	30,cycles
R5706	T19786	T19776	conj	cycles,step
R5707	T19787	T19786	prep	of,cycles
R5708	T19788	T19789	nummod	one,minute
R5709	T19789	T19790	compound	minute,denaturation
R571	T3565	T3563	parataxis	3,degeneration
R5710	T19790	T19787	pobj	denaturation,of
R5711	T19791	T19786	prep	at,cycles
R5712	T19792	T19793	nummod	95,°C
R5713	T19793	T19791	pobj	°C,at
R5714	T19794	T19786	punct	", ",cycles
R5715	T19795	T19796	nummod	one,minute
R5716	T19796	T19786	conj	minute,cycles
R5717	T19797	T19796	amod	annealing,minute
R5718	T19798	T19796	prep	at,minute
R5719	T19799	T19800	nummod	56,°C
R572	T3408	T3406	conj	distinct,specific
R5720	T19800	T19798	pobj	°C,at
R5721	T19801	T19796	punct	", ",minute
R5722	T19802	T19796	cc	and,minute
R5723	T19803	T19804	nummod	one,minute
R5724	T19804	T19805	compound	minute,extension
R5725	T19805	T19796	conj	extension,minute
R5726	T19806	T19805	prep	at,extension
R5727	T19807	T19808	nummod	72,°C
R5728	T19808	T19806	pobj	°C,at
R5729	T19809	T19776	punct	", ",step
R573	T3566	T3565	punct	),3
R5730	T19810	T19776	prep	with,step
R5731	T19811	T19812	det	a,step
R5732	T19812	T19810	pobj	step,with
R5733	T19813	T19814	nummod	ten,minute
R5734	T19814	T19812	nmod	minute,step
R5735	T19815	T19812	amod	final,step
R5736	T19816	T19812	compound	extension,step
R5737	T19817	T19774	punct	.,included
R5738	T19820	T19821	prep	Due,developed
R5739	T19822	T19820	pcomp	to,Due
R574	T3567	T3563	punct	", ",degeneration
R5740	T19823	T19824	det	the,amount
R5741	T19824	T19820	pobj	amount,Due
R5742	T19825	T19824	amod	limited,amount
R5743	T19826	T19824	prep	of,amount
R5744	T19827	T19828	amod	genetic,material
R5745	T19828	T19826	pobj	material,of
R5746	T19829	T19828	amod	available,material
R5747	T19830	T19824	prep	in,amount
R5748	T19831	T19832	amod	pre-implantation,embryos
R5749	T19832	T19830	pobj	embryos,in
R575	T3409	T3383	punct	.,suggest
R5750	T19833	T19821	punct	", ",developed
R5751	T19834	T19835	det	a,strategy
R5752	T19835	T19821	nsubjpass	strategy,developed
R5753	T19836	T19835	amod	nested,strategy
R5754	T19837	T19835	compound	PCR,strategy
R5755	T19838	T19821	auxpass	was,developed
R5756	T19839	T19840	aux	to,yield
R5757	T19840	T19821	advcl	yield,developed
R5758	T19841	T19842	amod	reliable,information
R5759	T19842	T19840	dobj	information,yield
R576	T3568	T3569	amod	neuronal,growth
R5760	T19843	T19842	compound	genotyping,information
R5761	T19844	T19845	punct	(,Figure
R5762	T19845	T19821	parataxis	Figure,developed
R5763	T19846	T19845	nummod	4,Figure
R5764	T19847	T19845	punct	),Figure
R5765	T19848	T19821	punct	.,developed
R5766	T19850	T19851	amod	Whole,embryos
R5767	T19851	T19852	nsubjpass	embryos,digested
R5768	T19853	T19852	auxpass	were,digested
R5769	T19854	T19852	advmod	first,digested
R577	T3569	T3563	conj	growth,degeneration
R5770	T19855	T19852	prep	in,digested
R5771	T19856	T19857	nummod	20,μL
R5772	T19857	T19855	pobj	μL,in
R5773	T19858	T19857	prep	of,μL
R5774	T19859	T19860	compound	proteinase,K
R5775	T19860	T19861	compound	K,buffer
R5776	T19861	T19858	pobj	buffer,of
R5777	T19862	T19863	punct	(,see
R5778	T19863	T19852	parataxis	see,digested
R5779	T19864	T19863	advmod	below,see
R578	T3411	T3412	det	The,designations
R5780	T19865	T19863	punct	),see
R5781	T19866	T19852	punct	.,digested
R5782	T19868	T19869	det	The,lysate
R5783	T19869	T19870	nsubjpass	lysate,divided
R5784	T19871	T19870	auxpass	was,divided
R5785	T19872	T19870	advmod	then,divided
R5786	T19873	T19870	prep	in,divided
R5787	T19874	T19873	pobj	two,in
R5788	T19875	T19870	punct	", ",divided
R5789	T19876	T19870	prep	with,divided
R579	T3570	T3569	cc	and,growth
R5790	T19877	T19878	nsubjpass	half,used
R5791	T19878	T19876	pcomp	used,with
R5792	T19879	T19880	punct	(,μL
R5793	T19880	T19877	parataxis	μL,half
R5794	T19881	T19880	nummod	10,μL
R5795	T19882	T19880	punct	),μL
R5796	T19883	T19878	auxpass	being,used
R5797	T19884	T19878	prep	in,used
R5798	T19885	T19886	det	the,amplification
R5799	T19886	T19884	pobj	amplification,in
R58	T730	T727	appos	calpain,Capn2
R580	T3571	T3569	conj	neurodegeneration,growth
R5800	T19887	T19886	prep	of,amplification
R5801	T19888	T19889	det	the,allele
R5802	T19889	T19887	pobj	allele,of
R5803	T19890	T19891	amod	wild,type
R5804	T19891	T19889	compound	type,allele
R5805	T19892	T19891	punct	-,type
R5806	T19893	T19878	cc	and,used
R5807	T19894	T19895	det	the,μL
R5808	T19895	T19878	conj	μL,used
R5809	T19896	T19895	amod	remaining,μL
R581	T3572	T3573	punct	[,4
R5810	T19897	T19895	nummod	10,μL
R5811	T19898	T19895	prep	in,μL
R5812	T19899	T19900	det	the,amplification
R5813	T19900	T19898	pobj	amplification,in
R5814	T19901	T19900	prep	of,amplification
R5815	T19902	T19903	det	the,allele
R5816	T19903	T19901	pobj	allele,of
R5817	T19904	T19903	compound	mutant,allele
R5818	T19905	T19870	punct	.,divided
R5819	T19907	T19908	det	The,reaction
R582	T3573	T3571	parataxis	4,neurodegeneration
R5820	T19908	T19910	nsubjpass	reaction,carried
R5821	T19909	T19908	amod	first,reaction
R5822	T19911	T19908	prep	in,reaction
R5823	T19912	T19913	det	the,amplification
R5824	T19913	T19911	pobj	amplification,in
R5825	T19914	T19913	amod	nested,amplification
R5826	T19915	T19913	compound	PCR,amplification
R5827	T19916	T19913	prep	of,amplification
R5828	T19917	T19918	det	the,allele
R5829	T19918	T19916	pobj	allele,of
R583	T3574	T3573	punct	],4
R5830	T19919	T19920	amod	wild,type
R5831	T19920	T19918	compound	type,allele
R5832	T19921	T19920	punct	-,type
R5833	T19922	T19910	auxpass	was,carried
R5834	T19923	T19910	prt	out,carried
R5835	T19924	T19910	prep	in,carried
R5836	T19925	T19926	det	a,volume
R5837	T19926	T19924	pobj	volume,in
R5838	T19927	T19926	amod	final,volume
R5839	T19928	T19926	compound	reaction,volume
R584	T3575	T3569	punct	", ",growth
R5840	T19929	T19926	prep	of,volume
R5841	T19930	T19931	nummod	50,μL
R5842	T19931	T19929	pobj	μL,of
R5843	T19932	T19910	punct	", ",carried
R5844	T19933	T19910	advcl	using,carried
R5845	T19934	T19935	det	an,primer
R5846	T19935	T19933	dobj	primer,using
R5847	T19936	T19935	nmod	intron,primer
R5848	T19937	T19936	nummod	6,intron
R5849	T19938	T19935	compound	sense,primer
R585	T3576	T3577	compound	cell,cycle
R5850	T19939	T19935	cc	and,primer
R5851	T19940	T19941	det	an,primer
R5852	T19941	T19935	conj	primer,primer
R5853	T19942	T19941	compound	antisense,primer
R5854	T19943	T19941	acl	located,primer
R5855	T19944	T19943	prep	in,located
R5856	T19945	T19944	pobj	intron,in
R5857	T19946	T19945	nummod	7,intron
R5858	T19947	T19910	punct	.,carried
R5859	T19949	T19950	nummod	Two,μL
R586	T3412	T3416	nsubj	designations,derive
R5860	T19950	T19951	nsubjpass	μL,used
R5861	T19952	T19950	prep	of,μL
R5862	T19953	T19954	det	the,reaction
R5863	T19954	T19952	pobj	reaction,of
R5864	T19955	T19954	amod	first,reaction
R5865	T19956	T19951	auxpass	were,used
R5866	T19957	T19951	prep	as,used
R5867	T19958	T19957	pobj	template,as
R5868	T19959	T19951	prep	in,used
R5869	T19960	T19961	det	the,amplification
R587	T3577	T3578	compound	cycle,progression
R5870	T19961	T19959	pobj	amplification,in
R5871	T19962	T19961	amod	second,amplification
R5872	T19963	T19961	compound	PCR,amplification
R5873	T19964	T19951	advcl	using,used
R5874	T19965	T19966	det	another,primer
R5875	T19966	T19964	dobj	primer,using
R5876	T19967	T19966	nmod	intron,primer
R5877	T19968	T19967	nummod	6,intron
R5878	T19969	T19966	compound	sense,primer
R5879	T19970	T19966	cc	and,primer
R588	T3578	T3569	conj	progression,growth
R5880	T19971	T19972	det	an,primer
R5881	T19972	T19966	conj	primer,primer
R5882	T19973	T19972	compound	antisense,primer
R5883	T19974	T19972	prep	in,primer
R5884	T19975	T19974	pobj	exon,in
R5885	T19976	T19975	nummod	7,exon
R5886	T19977	T19951	punct	.,used
R5887	T19979	T19980	det	The,amplification
R5888	T19980	T19982	nsubjpass	amplification,carried
R5889	T19981	T19980	amod	nested,amplification
R589	T3413	T3412	nmod	μ,designations
R5890	T19983	T19980	prep	of,amplification
R5891	T19984	T19985	det	the,allele
R5892	T19985	T19983	pobj	allele,of
R5893	T19986	T19985	compound	mutant,allele
R5894	T19987	T19982	auxpass	was,carried
R5895	T19988	T19982	prt	out,carried
R5896	T19989	T19982	advmod	similarly,carried
R5897	T19990	T19982	punct	.,carried
R5898	T19992	T19993	det	The,pair
R5899	T19993	T19996	nsubj	pair,consisted
R59	T731	T730	punct	-,calpain
R590	T3579	T3580	punct	[,6
R5900	T19994	T19993	amod	first,pair
R5901	T19995	T19993	compound	primer,pair
R5902	T19997	T19996	prep	of,consisted
R5903	T19998	T19999	det	the,primer
R5904	T19999	T19997	pobj	primer,of
R5905	T20000	T19999	nmod	intron,primer
R5906	T20001	T20000	nummod	6,intron
R5907	T20002	T19999	compound	sense,primer
R5908	T20003	T19999	cc	and,primer
R5909	T20004	T20005	det	an,primer
R591	T3580	T3578	parataxis	6,progression
R5910	T20005	T19999	conj	primer,primer
R5911	T20006	T20005	compound	antisense,primer
R5912	T20007	T20005	prep	in,primer
R5913	T20008	T20009	det	the,cassette
R5914	T20009	T20007	pobj	cassette,in
R5915	T20010	T20011	compound	PGK,Neo
R5916	T20011	T20009	compound	Neo,cassette
R5917	T20012	T20011	punct	-,Neo
R5918	T20013	T19996	punct	.,consisted
R5919	T20015	T20016	det	The,pair
R592	T3581	T3580	nummod	5,6
R5920	T20016	T20019	nsubjpass	pair,comprised
R5921	T20017	T20016	amod	nested,pair
R5922	T20018	T20016	compound	primer,pair
R5923	T20020	T20019	auxpass	was,comprised
R5924	T20021	T20019	prep	of,comprised
R5925	T20022	T20023	det	the,primer
R5926	T20023	T20021	pobj	primer,of
R5927	T20024	T20023	amod	second,primer
R5928	T20025	T20023	nmod	intron,primer
R5929	T20026	T20025	nummod	6,intron
R593	T3414	T3413	cc	and,μ
R5930	T20027	T20023	compound	sense,primer
R5931	T20028	T20023	cc	and,primer
R5932	T20029	T20030	det	an,primer
R5933	T20030	T20023	conj	primer,primer
R5934	T20031	T20030	compound	antisense,primer
R5935	T20032	T20033	advmod	also,located
R5936	T20033	T20030	acl	located,primer
R5937	T20034	T20033	prep	within,located
R5938	T20035	T20036	det	the,sequence
R5939	T20036	T20034	pobj	sequence,within
R594	T3582	T3580	punct	",",6
R5940	T20037	T20038	compound	PGK,Neo
R5941	T20038	T20036	compound	Neo,sequence
R5942	T20039	T20038	punct	-,Neo
R5943	T20040	T20019	punct	.,comprised
R5944	T20042	T20043	nsubjpass	It,noted
R5945	T20044	T20043	aux	should,noted
R5946	T20045	T20043	auxpass	be,noted
R5947	T20046	T20047	mark	that,involved
R5948	T20047	T20043	ccomp	involved,noted
R5949	T20048	T20047	nsubj	amplification,involved
R595	T3583	T3580	punct	],6
R5950	T20049	T20048	prep	of,amplification
R5951	T20050	T20051	det	both,sequences
R5952	T20051	T20049	pobj	sequences,of
R5953	T20052	T20053	det	the,primers
R5954	T20053	T20047	dobj	primers,involved
R5955	T20054	T20053	amod	same,primers
R5956	T20055	T20053	compound	sense,primers
R5957	T20056	T20047	prep	in,involved
R5958	T20057	T20058	det	both,steps
R5959	T20058	T20056	pobj	steps,in
R596	T3584	T3578	punct	", ",progression
R5960	T20059	T20058	prep	of,steps
R5961	T20060	T20061	det	the,strategy
R5962	T20061	T20059	pobj	strategy,of
R5963	T20062	T20061	amod	nested,strategy
R5964	T20063	T20061	compound	PCR,strategy
R5965	T20064	T20043	punct	.,noted
R5966	T20066	T20067	prep	In,included
R5967	T20068	T20066	pobj	addition,In
R5968	T20069	T20067	punct	", ",included
R5969	T20070	T20071	det	the,sets
R597	T3415	T3413	conj	m,μ
R5970	T20071	T20067	nsubj	sets,included
R5971	T20072	T20071	nummod	two,sets
R5972	T20073	T20071	prep	of,sets
R5973	T20074	T20073	pobj	reactions,of
R5974	T20075	T20076	det	a,control
R5975	T20076	T20067	dobj	control,included
R5976	T20077	T20076	amod	common,control
R5977	T20078	T20076	amod	internal,control
R5978	T20079	T20076	acl	designed,control
R5979	T20080	T20081	aux	to,amplify
R598	T3585	T3586	compound	signal,cascades
R5980	T20081	T20079	advcl	amplify,designed
R5981	T20082	T20083	det	a,region
R5982	T20083	T20081	dobj	region,amplify
R5983	T20084	T20083	amod	genomic,region
R5984	T20085	T20083	prep	within,region
R5985	T20086	T20087	det	the,arm
R5986	T20087	T20085	pobj	arm,within
R5987	T20088	T20087	amod	short,arm
R5988	T20089	T20087	prep	of,arm
R5989	T20090	T20091	det	the,vector
R599	T3586	T3578	conj	cascades,progression
R5990	T20091	T20089	pobj	vector,of
R5991	T20092	T20091	compound	targeting,vector
R5992	T20093	T20094	dep	that,preserved
R5993	T20094	T20083	relcl	preserved,region
R5994	T20095	T20094	auxpass	is,preserved
R5995	T20096	T20094	prep	in,preserved
R5996	T20097	T20098	preconj	both,alleles
R5997	T20098	T20096	pobj	alleles,in
R5998	T20099	T20098	det	the,alleles
R5999	T20100	T20101	amod	wild,type
R6	T670	T669	auxpass	is,required
R60	T732	T715	punct	),subunits
R600	T3587	T3586	acl	triggered,cascades
R6000	T20101	T20098	nmod	type,alleles
R6001	T20102	T20101	punct	-,type
R6002	T20103	T20101	cc	and,type
R6003	T20104	T20101	conj	mutant,type
R6004	T20105	T20067	punct	.,included
R6005	T20107	T20108	det	The,pair
R6006	T20108	T20111	nsubjpass	pair,made
R6007	T20109	T20108	amod	first,pair
R6008	T20110	T20108	compound	primer,pair
R6009	T20112	T20108	prep	of,pair
R601	T3417	T3416	prep	from,derive
R6010	T20113	T20114	det	the,PCR
R6011	T20114	T20112	pobj	PCR,of
R6012	T20115	T20114	compound	control,PCR
R6013	T20116	T20111	auxpass	was,made
R6014	T20117	T20111	prt	up,made
R6015	T20118	T20111	prep	of,made
R6016	T20119	T20120	det	an,primer
R6017	T20120	T20118	pobj	primer,of
R6018	T20121	T20120	nmod	intron,primer
R6019	T20122	T20121	nummod	4,intron
R602	T3588	T3587	agent	by,triggered
R6020	T20123	T20120	compound	sense,primer
R6021	T20124	T20120	cc	and,primer
R6022	T20125	T20126	det	an,primer
R6023	T20126	T20120	conj	primer,primer
R6024	T20127	T20126	nmod	intron,primer
R6025	T20128	T20127	nummod	5,intron
R6026	T20129	T20126	compound	antisense,primer
R6027	T20130	T20111	punct	.,made
R6028	T20132	T20133	det	The,pair
R6029	T20133	T20136	nsubj	pair,comprised
R603	T3589	T3588	pobj	integrins,by
R6030	T20134	T20133	amod	second,pair
R6031	T20135	T20133	compound	primer,pair
R6032	T20137	T20138	det	an,primer
R6033	T20138	T20136	dobj	primer,comprised
R6034	T20139	T20138	nmod	exon,primer
R6035	T20140	T20139	nummod	5,exon
R6036	T20141	T20138	compound	sense,primer
R6037	T20142	T20138	cc	and,primer
R6038	T20143	T20144	det	a,primer
R6039	T20144	T20138	conj	primer,primer
R604	T3590	T3589	cc	and,integrins
R6040	T20145	T20144	amod	nested,primer
R6041	T20146	T20144	nmod	intron,primer
R6042	T20147	T20146	nummod	5,intron
R6043	T20148	T20144	compound	antisense,primer
R6044	T20149	T20136	punct	.,comprised
R6045	T20151	T20152	det	The,products
R6046	T20152	T20154	nsubj	products,were
R6047	T20153	T20152	amod	final,products
R6048	T20155	T20156	nummod	213,bp
R6049	T20156	T20154	attr	bp,were
R605	T3591	T3592	compound	growth,factors
R6050	T20157	T20154	prep	for,were
R6051	T20158	T20159	det	the,PCR
R6052	T20159	T20157	pobj	PCR,for
R6053	T20160	T20159	compound	control,PCR
R6054	T20161	T20154	punct	", ",were
R6055	T20162	T20163	nummod	429,bp
R6056	T20163	T20154	conj	bp,were
R6057	T20164	T20163	prep	for,bp
R6058	T20165	T20166	det	the,PCR
R6059	T20166	T20164	pobj	PCR,for
R606	T3418	T3419	det	the,levels
R6060	T20167	T20168	amod	wild,type
R6061	T20168	T20166	compound	type,PCR
R6062	T20169	T20168	punct	-,type
R6063	T20170	T20163	punct	", ",bp
R6064	T20171	T20163	cc	and,bp
R6065	T20172	T20173	nummod	389,bp
R6066	T20173	T20163	conj	bp,bp
R6067	T20174	T20173	prep	for,bp
R6068	T20175	T20176	det	the,PCR
R6069	T20176	T20174	pobj	PCR,for
R607	T3592	T3589	conj	factors,integrins
R6070	T20177	T20176	compound	mutant,PCR
R6071	T20178	T20154	punct	.,were
R6072	T20180	T20181	det	All,reactions
R6073	T20181	T20182	nsubjpass	reactions,carried
R6074	T20183	T20182	auxpass	were,carried
R6075	T20184	T20182	prt	out,carried
R6076	T20185	T20182	advcl	using,carried
R6077	T20186	T20187	amod	identical,conditions
R6078	T20187	T20185	dobj	conditions,using
R6079	T20188	T20187	compound	PCR,conditions
R608	T3593	T3594	punct	[,7
R6080	T20189	T20187	acl	entailing,conditions
R6081	T20190	T20191	det	an,denaturation
R6082	T20191	T20189	dobj	denaturation,entailing
R6083	T20192	T20191	amod	initial,denaturation
R6084	T20193	T20194	nummod	five,minute
R6085	T20194	T20191	compound	minute,denaturation
R6086	T20195	T20191	prep	at,denaturation
R6087	T20196	T20197	nummod	95,°C
R6088	T20197	T20195	pobj	°C,at
R6089	T20198	T20191	punct	", ",denaturation
R609	T3594	T3587	parataxis	7,triggered
R6090	T20199	T20200	nummod	35,cycles
R6091	T20200	T20191	conj	cycles,denaturation
R6092	T20201	T20200	prep	of,cycles
R6093	T20202	T20203	nummod	one,denaturation
R6094	T20203	T20201	pobj	denaturation,of
R6095	T20204	T20203	compound	minute,denaturation
R6096	T20205	T20200	prep	at,cycles
R6097	T20206	T20207	nummod	95,°C
R6098	T20207	T20205	pobj	°C,at
R6099	T20208	T20200	punct	", ",cycles
R61	T733	T715	punct	", ",subunits
R610	T3419	T3417	pobj	levels,from
R6100	T20209	T20210	nummod	one,minute
R6101	T20210	T20200	conj	minute,cycles
R6102	T20211	T20210	amod	annealing,minute
R6103	T20212	T20210	prep	at,minute
R6104	T20213	T20214	nummod	56,°C
R6105	T20214	T20212	pobj	°C,at
R6106	T20215	T20210	punct	", ",minute
R6107	T20216	T20210	cc	and,minute
R6108	T20217	T20218	nummod	one,minute
R6109	T20218	T20219	compound	minute,extension
R611	T3595	T3594	punct	],7
R6110	T20219	T20210	conj	extension,minute
R6111	T20220	T20219	prep	at,extension
R6112	T20221	T20222	nummod	72,°C
R6113	T20222	T20220	pobj	°C,at
R6114	T20223	T20189	punct	", ",entailing
R6115	T20224	T20189	prep	with,entailing
R6116	T20225	T20226	det	a,step
R6117	T20226	T20224	pobj	step,with
R6118	T20227	T20226	amod	final,step
R6119	T20228	T20226	compound	extension,step
R612	T3596	T3586	punct	", ",cascades
R6120	T20229	T20226	prep	of,step
R6121	T20230	T20231	nummod	ten,minutes
R6122	T20231	T20229	pobj	minutes,of
R6123	T20232	T20182	punct	.,carried
R6124	T20493	T20492	prep	of,Isolation
R6125	T20494	T20495	amod	pre-implantation,embryos
R6126	T20495	T20493	pobj	embryos,of
R6127	T20497	T20498	nsubjpass	Time,determined
R6128	T20499	T20497	prep	of,Time
R6129	T20500	T20499	pobj	fertilization,of
R613	T3597	T3598	compound	membrane,protrusion
R6130	T20501	T20498	auxpass	was,determined
R6131	T20502	T20498	agent	by,determined
R6132	T20503	T20502	pobj	observation,by
R6133	T20504	T20503	prep	of,observation
R6134	T20505	T20506	compound	copulation,plugs
R6135	T20506	T20504	pobj	plugs,of
R6136	T20507	T20498	punct	", ",determined
R6137	T20508	T20498	cc	and,determined
R6138	T20509	T20510	nsubjpass	noon,defined
R6139	T20510	T20498	conj	defined,determined
R614	T3598	T3586	conj	protrusion,cascades
R6140	T20511	T20509	prep	of,noon
R6141	T20512	T20513	det	that,day
R6142	T20513	T20511	pobj	day,of
R6143	T20514	T20510	auxpass	was,defined
R6144	T20515	T20510	prep	as,defined
R6145	T20516	T20515	pobj	E0.5,as
R6146	T20517	T20510	punct	.,defined
R6147	T20519	T20520	amod	Pre-implantation,embryos
R6148	T20520	T20521	nsubjpass	embryos,obtained
R6149	T20522	T20521	auxpass	were,obtained
R615	T3420	T3419	prep	of,levels
R6150	T20523	T20521	prep	by,obtained
R6151	T20524	T20523	pcomp	dissecting,by
R6152	T20525	T20526	det	the,uteri
R6153	T20526	T20524	dobj	uteri,dissecting
R6154	T20527	T20524	prep	out,dissecting
R6155	T20528	T20527	prep	of,out
R6156	T20529	T20530	amod	pregnant,females
R6157	T20530	T20528	pobj	females,of
R6158	T20531	T20524	prep	at,dissecting
R6159	T20532	T20531	pobj	E2.5,at
R616	T3599	T3600	punct	[,8
R6160	T20533	T20532	cc	or,E2.5
R6161	T20534	T20532	conj	E3.5,E2.5
R6162	T20535	T20524	cc	and,dissecting
R6163	T20536	T20524	conj	flushing,dissecting
R6164	T20537	T20538	det	the,oviducts
R6165	T20538	T20536	dobj	oviducts,flushing
R6166	T20539	T20536	prep	with,flushing
R6167	T20540	T20541	compound	ES,medium
R6168	T20541	T20539	pobj	medium,with
R6169	T20542	T20541	compound	cell,medium
R617	T3600	T3598	parataxis	8,protrusion
R6170	T20543	T20521	punct	.,obtained
R6171	T20545	T20546	amod	Isolated,embryos
R6172	T20546	T20547	nsubjpass	embryos,digested
R6173	T20548	T20547	auxpass	were,digested
R6174	T20549	T20547	advmod	then,digested
R6175	T20550	T20547	prep	for,digested
R6176	T20551	T20552	nummod	five,hours
R6177	T20552	T20550	pobj	hours,for
R6178	T20553	T20547	prep	at,digested
R6179	T20554	T20555	nummod	55,°C
R618	T3421	T3420	pobj	Ca2,of
R6180	T20555	T20553	pobj	°C,at
R6181	T20556	T20547	prep	in,digested
R6182	T20557	T20558	nummod	20,μL
R6183	T20558	T20556	pobj	μL,in
R6184	T20559	T20558	prep	of,μL
R6185	T20560	T20561	compound	lysis,buffer
R6186	T20561	T20559	pobj	buffer,of
R6187	T20562	T20563	punct	(,pH
R6188	T20563	T20561	parataxis	pH,buffer
R6189	T20564	T20565	nummod	50,mM
R619	T3601	T3600	punct	],8
R6190	T20565	T20566	compound	mM,HCl
R6191	T20566	T20563	dep	HCl,pH
R6192	T20567	T20566	compound	Tris,HCl
R6193	T20568	T20566	punct	-,HCl
R6194	T20569	T20563	punct	", ",pH
R6195	T20570	T20571	nummod	0.5,%
R6196	T20571	T20572	compound	%,X
R6197	T20572	T20563	dep	X,pH
R6198	T20573	T20572	compound	Triton,X
R6199	T20574	T20572	punct	-,X
R62	T734	T715	cc	and,subunits
R620	T3602	T3598	punct	", ",protrusion
R6200	T20575	T20572	nummod	100,X
R6201	T20576	T20563	punct	", ",pH
R6202	T20577	T20578	nummod	200,μL
R6203	T20578	T20579	nmod	μL,K
R6204	T20579	T20563	dep	K,pH
R6205	T20580	T20581	punct	/,mL
R6206	T20581	T20578	prep	mL,μL
R6207	T20582	T20579	compound	proteinase,K
R6208	T20583	T20563	punct	", ",pH
R6209	T20584	T20563	nummod	8.0,pH
R621	T3603	T3598	conj	remodeling,protrusion
R6210	T20585	T20563	punct	),pH
R6211	T20586	T20561	punct	", ",buffer
R6212	T20587	T20561	acl	followed,buffer
R6213	T20588	T20587	agent	by,followed
R6214	T20589	T20590	nummod	ten,minutes
R6215	T20590	T20588	pobj	minutes,by
R6216	T20591	T20590	prep	at,minutes
R6217	T20592	T20593	nummod	95,°C
R6218	T20593	T20591	pobj	°C,at
R6219	T20594	T20595	aux	to,inactivate
R622	T3422	T3421	punct	+,Ca2
R6220	T20595	T20587	advcl	inactivate,followed
R6221	T20596	T20597	det	the,K
R6222	T20597	T20595	dobj	K,inactivate
R6223	T20598	T20597	compound	proteinase,K
R6224	T20599	T20547	punct	.,digested
R6225	T20601	T20602	nsubjpass	Lysates,used
R6226	T20603	T20602	auxpass	were,used
R6227	T20604	T20602	advmod	then,used
R6228	T20605	T20602	prep	for,used
R6229	T20606	T20607	compound	PCR,genotyping
R623	T3604	T3603	prep	of,remodeling
R6230	T20607	T20605	pobj	genotyping,for
R6231	T20608	T20602	punct	.,used
R6232	T21152	T21151	prep	of,Characterization
R6233	T21153	T21154	amod	targeted,lines
R6234	T21154	T21152	pobj	lines,of
R6235	T21155	T21156	compound	ES,cell
R6236	T21156	T21154	compound	cell,lines
R6237	T21157	T21151	punct	.,Characterization
R6238	T21159	T21160	punct	(,A
R6239	T21160	T21161	meta	A,detected
R624	T3605	T3606	det	the,cytoskeleton
R6240	T21162	T21160	punct	),A
R6241	T21163	T21164	amod	Targeted,disruption
R6242	T21164	T21161	nsubjpass	disruption,detected
R6243	T21165	T21164	prep	of,disruption
R6244	T21166	T21167	det	the,locus
R6245	T21167	T21165	pobj	locus,of
R6246	T21168	T21167	compound	Capn2,locus
R6247	T21169	T21161	auxpass	was,detected
R6248	T21170	T21161	advmod	initially,detected
R6249	T21171	T21161	agent	by,detected
R625	T3606	T3604	pobj	cytoskeleton,of
R6250	T21172	T21173	compound	Southern,blotting
R6251	T21173	T21171	pobj	blotting,by
R6252	T21174	T21161	punct	.,detected
R6253	T21176	T21177	nsubjpass	Membranes,blotted
R6254	T21178	T21177	auxpass	were,blotted
R6255	T21179	T21177	prep	with,blotted
R6256	T21180	T21181	npadvmod	BamHI,digested
R6257	T21181	T21183	amod	digested,DNA
R6258	T21182	T21181	punct	-,digested
R6259	T21183	T21179	pobj	DNA,with
R626	T3423	T3419	acl	required,levels
R6260	T21184	T21183	amod	genomic,DNA
R6261	T21185	T21183	acl	extracted,DNA
R6262	T21186	T21185	prep	from,extracted
R6263	T21187	T21188	compound	ES,cells
R6264	T21188	T21186	pobj	cells,from
R6265	T21189	T21177	cc	and,blotted
R6266	T21190	T21177	conj	hybridized,blotted
R6267	T21191	T21190	prep	with,hybridized
R6268	T21192	T21193	det	a,fragment
R6269	T21193	T21191	pobj	fragment,with
R627	T3607	T3603	cc	and,remodeling
R6270	T21194	T21195	npadvmod	DIG,labeled
R6271	T21195	T21193	amod	labeled,fragment
R6272	T21196	T21195	punct	-,labeled
R6273	T21197	T21198	nummod	823,bp
R6274	T21198	T21193	compound	bp,fragment
R6275	T21199	T21200	compound	BamHI,HindIII
R6276	T21200	T21193	compound	HindIII,fragment
R6277	T21201	T21200	punct	/,HindIII
R6278	T21202	T21193	acl	located,fragment
R6279	T21203	T21202	advmod	immediately,located
R628	T3608	T3609	compound	cell,migration
R6280	T21204	T21203	advmod	upstream,immediately
R6281	T21205	T21203	prep	of,immediately
R6282	T21206	T21207	det	the,arm
R6283	T21207	T21205	pobj	arm,of
R6284	T21208	T21207	amod	short,arm
R6285	T21209	T21207	prep	of,arm
R6286	T21210	T21211	det	the,vector
R6287	T21211	T21209	pobj	vector,of
R6288	T21212	T21211	compound	targeting,vector
R6289	T21213	T21214	punct	(,Figure
R629	T3609	T3603	conj	migration,remodeling
R6290	T21214	T21190	parataxis	Figure,hybridized
R6291	T21215	T21214	nummod	1,Figure
R6292	T21216	T21214	punct	),Figure
R6293	T21217	T21177	punct	.,blotted
R6294	T21219	T21220	det	A,fragment
R6295	T21220	T21225	nsubj	fragment,was
R6296	T21221	T21222	nummod	3.5,kb
R6297	T21222	T21220	compound	kb,fragment
R6298	T21223	T21222	punct	-,kb
R6299	T21224	T21220	compound	BamHI,fragment
R63	T735	T736	det	a,subunit
R630	T3424	T3425	advmod	in,vitro
R6300	T21226	T21220	acl	corresponding,fragment
R6301	T21227	T21226	prep	to,corresponding
R6302	T21228	T21229	det	the,allele
R6303	T21229	T21227	pobj	allele,to
R6304	T21230	T21231	amod	wild,type
R6305	T21231	T21229	compound	type,allele
R6306	T21232	T21231	punct	-,type
R6307	T21233	T21225	acomp	present,was
R6308	T21234	T21225	prep	in,was
R6309	T21235	T21236	det	all,cells
R631	T3610	T3611	punct	[,9
R6310	T21236	T21234	pobj	cells,in
R6311	T21237	T21225	punct	", ",was
R6312	T21238	T21239	mark	whereas,detected
R6313	T21239	T21225	advcl	detected,was
R6314	T21240	T21241	det	a,fragment
R6315	T21241	T21239	nsubjpass	fragment,detected
R6316	T21242	T21243	nummod	5.3,kb
R6317	T21243	T21241	compound	kb,fragment
R6318	T21244	T21243	punct	-,kb
R6319	T21245	T21241	prep	from,fragment
R632	T3611	T3609	parataxis	9,migration
R6320	T21246	T21247	det	the,allele
R6321	T21247	T21245	pobj	allele,from
R6322	T21248	T21247	compound	mutant,allele
R6323	T21249	T21239	auxpass	was,detected
R6324	T21250	T21239	prep	in,detected
R6325	T21251	T21252	nummod	two,lines
R6326	T21252	T21250	pobj	lines,in
R6327	T21253	T21252	amod	targeted,lines
R6328	T21254	T21252	compound	cell,lines
R6329	T21255	T21239	punct	", ",detected
R633	T3612	T3613	punct	-,15
R6330	T21256	T21239	dep	designated,detected
R6331	T21257	T21256	oprd	ES27,designated
R6332	T21258	T21257	cc	and,ES27
R6333	T21259	T21257	conj	ES36,ES27
R6334	T21260	T21225	punct	.,was
R6335	T21262	T21263	punct	(,B
R6336	T21263	T21264	meta	B,carried
R6337	T21265	T21263	punct	),B
R6338	T21266	T21267	compound	PCR,genotyping
R6339	T21267	T21264	nsubjpass	genotyping,carried
R634	T3613	T3611	appos	15,9
R6340	T21268	T21264	auxpass	was,carried
R6341	T21269	T21264	prt	out,carried
R6342	T21270	T21264	prep	with,carried
R6343	T21271	T21272	nummod	two,reactions
R6344	T21272	T21270	pobj	reactions,with
R6345	T21273	T21272	amod	separate,reactions
R6346	T21274	T21272	acl	designed,reactions
R6347	T21275	T21276	aux	to,amplify
R6348	T21276	T21274	advcl	amplify,designed
R6349	T21277	T21278	preconj	either,segment
R635	T3614	T3611	punct	],9
R6350	T21278	T21276	dobj	segment,amplify
R6351	T21279	T21278	det	a,segment
R6352	T21280	T21281	nummod	"2,748",bp
R6353	T21281	T21278	compound	bp,segment
R6354	T21282	T21278	prep	from,segment
R6355	T21283	T21284	det	the,allele
R6356	T21284	T21282	pobj	allele,from
R6357	T21285	T21286	amod	wild,type
R6358	T21286	T21284	compound	type,allele
R6359	T21287	T21286	punct	-,type
R636	T3615	T3603	punct	", ",remodeling
R6360	T21288	T21278	cc	or,segment
R6361	T21289	T21290	det	a,segment
R6362	T21290	T21278	conj	segment,segment
R6363	T21291	T21292	nummod	"2,711",bp
R6364	T21292	T21290	compound	bp,segment
R6365	T21293	T21290	prep	from,segment
R6366	T21294	T21295	det	the,allele
R6367	T21295	T21293	pobj	allele,from
R6368	T21296	T21295	compound	mutant,allele
R6369	T21297	T21264	punct	.,carried
R637	T3425	T3423	advmod	vitro,required
R6370	T21299	T21300	det	Both,reactions
R6371	T21300	T21301	nsubj	reactions,used
R6372	T21302	T21303	det	a,primer
R6373	T21303	T21301	dobj	primer,used
R6374	T21304	T21305	amod	common,sense
R6375	T21305	T21303	compound	sense,primer
R6376	T21306	T21303	acl	located,primer
R6377	T21307	T21306	prep	in,located
R6378	T21308	T21307	pobj	intron,in
R6379	T21309	T21308	nummod	4,intron
R638	T3616	T3603	cc	and,remodeling
R6380	T21310	T21306	punct	", ",located
R6381	T21311	T21306	prep	outside,located
R6382	T21312	T21313	det	the,arm
R6383	T21313	T21311	pobj	arm,outside
R6384	T21314	T21313	amod	short,arm
R6385	T21315	T21313	prep	of,arm
R6386	T21316	T21317	det	the,vector
R6387	T21317	T21315	pobj	vector,of
R6388	T21318	T21317	compound	targeting,vector
R6389	T21319	T21313	punct	", ",arm
R639	T3617	T3603	conj	regulation,remodeling
R6390	T21320	T21313	cc	and,arm
R6391	T21321	T21322	amod	distinct,primers
R6392	T21322	T21313	conj	primers,arm
R6393	T21323	T21324	npadvmod	allele,specific
R6394	T21324	T21322	amod	specific,primers
R6395	T21325	T21324	punct	-,specific
R6396	T21326	T21322	compound	antisense,primers
R6397	T21327	T21301	punct	.,used
R6398	T21329	T21330	det	The,reaction
R6399	T21330	T21331	nsubj	reaction,used
R64	T736	T715	conj	subunit,subunits
R640	T3426	T3423	prep	for,required
R6400	T21332	T21333	aux	to,detect
R6401	T21333	T21330	advcl	detect,reaction
R6402	T21334	T21335	det	the,allele
R6403	T21335	T21333	dobj	allele,detect
R6404	T21336	T21337	amod	wild,type
R6405	T21337	T21335	compound	type,allele
R6406	T21338	T21337	punct	-,type
R6407	T21339	T21340	det	an,primer
R6408	T21340	T21331	dobj	primer,used
R6409	T21341	T21340	compound	antisense,primer
R641	T3618	T3617	prep	of,regulation
R6410	T21342	T21340	acl	located,primer
R6411	T21343	T21342	prep	in,located
R6412	T21344	T21343	pobj	exon,in
R6413	T21345	T21344	nummod	7,exon
R6414	T21346	T21347	mark	while,done
R6415	T21347	T21331	advcl	done,used
R6416	T21348	T21349	det	the,amplification
R6417	T21349	T21347	nsubjpass	amplification,done
R6418	T21350	T21349	prep	of,amplification
R6419	T21351	T21352	det	the,sequence
R642	T3619	T3620	compound	cell,death
R6420	T21352	T21350	pobj	sequence,of
R6421	T21353	T21352	compound	mutant,sequence
R6422	T21354	T21347	auxpass	was,done
R6423	T21355	T21347	prep	with,done
R6424	T21356	T21357	det	an,primer
R6425	T21357	T21355	pobj	primer,with
R6426	T21358	T21357	compound	antisense,primer
R6427	T21359	T21357	prep	in,primer
R6428	T21360	T21361	det	the,cassette
R6429	T21361	T21359	pobj	cassette,in
R643	T3620	T3618	pobj	death,of
R6430	T21362	T21363	compound	PGK,Neo
R6431	T21363	T21361	compound	Neo,cassette
R6432	T21364	T21363	punct	-,Neo
R6433	T21365	T21331	punct	.,used
R6434	T21367	T21368	det	The,results
R6435	T21368	T21369	nsubj	results,confirm
R6436	T21370	T21371	det	the,presence
R6437	T21371	T21369	dobj	presence,confirm
R6438	T21372	T21371	prep	of,presence
R6439	T21373	T21374	det	the,allele
R644	T3621	T3617	prep	via,regulation
R6440	T21374	T21372	pobj	allele,of
R6441	T21375	T21376	amod	wild,type
R6442	T21376	T21374	compound	type,allele
R6443	T21377	T21376	punct	-,type
R6444	T21378	T21371	prep	in,presence
R6445	T21379	T21380	det	all,cells
R6446	T21380	T21378	pobj	cells,in
R6447	T21381	T21369	punct	", ",confirm
R6448	T21382	T21383	mark	whereas,observed
R6449	T21383	T21369	advcl	observed,confirm
R645	T3427	T3428	amod	optimal,activation
R6450	T21384	T21385	det	the,signal
R6451	T21385	T21383	nsubjpass	signal,observed
R6452	T21386	T21385	compound	mutant,signal
R6453	T21387	T21385	compound	allele,signal
R6454	T21388	T21383	auxpass	was,observed
R6455	T21389	T21390	advmod	only,in
R6456	T21390	T21383	prep	in,observed
R6457	T21391	T21392	det	the,clones
R6458	T21392	T21390	pobj	clones,in
R6459	T21393	T21392	nummod	two,clones
R646	T3622	T3623	preconj	both,necrosis
R6460	T21394	T21392	amod	targeted,clones
R6461	T21395	T21369	punct	.,confirm
R6462	T21397	T21398	punct	(,denotes
R6463	T21399	T21398	nsubj	M,denotes
R6464	T21400	T21398	punct	),denotes
R6465	T21401	T21402	det	the,marker
R6466	T21402	T21398	dobj	marker,denotes
R6467	T21403	T21404	amod	molecular,weight
R6468	T21404	T21402	compound	weight,marker
R6469	T21405	T21398	punct	.,denotes
R647	T3623	T3621	pobj	necrosis,via
R6470	T21857	T21858	amod	Nested,strategy
R6471	T21859	T21858	compound	PCR,strategy
R6472	T21860	T21858	prep	for,strategy
R6473	T21861	T21860	pobj	genotyping,for
R6474	T21862	T21861	prep	of,genotyping
R6475	T21863	T21864	amod	pre-implantation,embryos
R6476	T21864	T21862	pobj	embryos,of
R6477	T21865	T21858	punct	.,strategy
R6478	T21867	T21868	prep	Due,developed
R6479	T21869	T21867	pcomp	to,Due
R648	T3624	T3623	cc	and,necrosis
R6480	T21870	T21871	det	the,scarcity
R6481	T21871	T21867	pobj	scarcity,Due
R6482	T21872	T21871	prep	of,scarcity
R6483	T21873	T21874	amod	extractable,material
R6484	T21874	T21872	pobj	material,of
R6485	T21875	T21874	amod	genetic,material
R6486	T21876	T21868	punct	", ",developed
R6487	T21877	T21878	det	a,strategy
R6488	T21878	T21868	nsubjpass	strategy,developed
R6489	T21879	T21878	amod	nested,strategy
R649	T3428	T3426	pobj	activation,for
R6490	T21880	T21878	compound	PCR,strategy
R6491	T21881	T21868	auxpass	was,developed
R6492	T21882	T21868	prep	in,developed
R6493	T21883	T21882	pobj	order,in
R6494	T21884	T21885	aux	to,genotype
R6495	T21885	T21883	acl	genotype,order
R6496	T21886	T21887	amod	pre-implantation,embryos
R6497	T21887	T21885	dobj	embryos,genotype
R6498	T21888	T21868	punct	.,developed
R6499	T21890	T21891	amod	Separate,reactions
R65	T737	T736	amod	common,subunit
R650	T3625	T3623	conj	apoptosis,necrosis
R6500	T21891	T21892	nsubjpass	reactions,used
R6501	T21893	T21892	auxpass	were,used
R6502	T21894	T21895	aux	to,amplify
R6503	T21895	T21892	advcl	amplify,used
R6504	T21896	T21897	det	a,fragment
R6505	T21897	T21895	dobj	fragment,amplify
R6506	T21898	T21899	nummod	429,bp
R6507	T21899	T21897	compound	bp,fragment
R6508	T21900	T21897	prep	from,fragment
R6509	T21901	T21902	det	the,allele
R651	T3626	T3627	punct	[,16
R6510	T21902	T21900	pobj	allele,from
R6511	T21903	T21904	amod	wild,type
R6512	T21904	T21902	compound	type,allele
R6513	T21905	T21904	punct	-,type
R6514	T21906	T21897	cc	and,fragment
R6515	T21907	T21908	det	a,segment
R6516	T21908	T21897	conj	segment,fragment
R6517	T21909	T21910	nummod	389,bp
R6518	T21910	T21908	compound	bp,segment
R6519	T21911	T21908	prep	from,segment
R652	T3627	T3617	parataxis	16,regulation
R6520	T21912	T21913	det	the,allele
R6521	T21913	T21911	pobj	allele,from
R6522	T21914	T21913	compound	mutant,allele
R6523	T21915	T21897	punct	", ",fragment
R6524	T21916	T21897	appos	both,fragment
R6525	T21917	T21916	acl	spanning,both
R6526	T21918	T21919	det	the,end
R6527	T21919	T21917	dobj	end,spanning
R6528	T21920	T21919	nummod	3,end
R6529	T21921	T21920	punct	',3
R653	T3640	T3634	dobj	interplay,suggests
R6530	T21922	T21919	prep	of,end
R6531	T21923	T21924	det	the,arm
R6532	T21924	T21922	pobj	arm,of
R6533	T21925	T21924	amod	short,arm
R6534	T21926	T21927	punct	(,upstream
R6535	T21927	T21924	parataxis	upstream,arm
R6536	T21928	T21927	punct	),upstream
R6537	T21929	T21924	prep	of,arm
R6538	T21930	T21931	det	the,vector
R6539	T21931	T21929	pobj	vector,of
R654	T3628	T3629	punct	-,22
R6540	T21932	T21931	compound	targeting,vector
R6541	T21933	T21892	punct	.,used
R6542	T21935	T21936	prep	In,co-amplified
R6543	T21937	T21938	det	both,reactions
R6544	T21938	T21935	pobj	reactions,In
R6545	T21939	T21936	punct	", ",co-amplified
R6546	T21940	T21941	det	a,sequence
R6547	T21941	T21936	nsubjpass	sequence,co-amplified
R6548	T21942	T21943	nummod	213,bp
R6549	T21943	T21941	compound	bp,sequence
R655	T3629	T3627	appos	22,16
R6550	T21944	T21941	acl	located,sequence
R6551	T21945	T21944	prep	within,located
R6552	T21946	T21947	det	the,arm
R6553	T21947	T21945	pobj	arm,within
R6554	T21948	T21947	amod	short,arm
R6555	T21949	T21936	auxpass	was,co-amplified
R6556	T21950	T21936	prep	with,co-amplified
R6557	T21951	T21952	det	the,products
R6558	T21952	T21950	pobj	products,with
R6559	T21953	T21952	punct	',products
R656	T3630	T3627	punct	],16
R6560	T21954	T21952	amod	diagnostic,products
R6561	T21955	T21952	punct	',products
R6562	T21956	T21936	prep	as,co-amplified
R6563	T21957	T21958	det	an,control
R6564	T21958	T21956	pobj	control,as
R6565	T21959	T21958	amod	internal,control
R6566	T21960	T21936	punct	.,co-amplified
R6567	T21962	T21963	det	The,primers
R6568	T21963	T21966	nsubjpass	primers,used
R6569	T21964	T21963	amod	same,primers
R657	T3631	T3546	punct	.,linked
R6570	T21965	T21963	compound	sense,primers
R6571	T21967	T21966	auxpass	were,used
R6572	T21968	T21969	aux	to,amplify
R6573	T21969	T21966	advcl	amplify,used
R6574	T21970	T21969	punct	',amplify
R6575	T21971	T21972	amod	diagnostic,sequences
R6576	T21972	T21969	dobj	sequences,amplify
R6577	T21973	T21972	punct	',sequences
R6578	T21974	T21972	prep	in,sequences
R6579	T21975	T21976	det	both,reactions
R658	T3641	T3640	amod	complex,interplay
R6580	T21976	T21974	pobj	reactions,in
R6581	T21977	T21969	punct	", ",amplify
R6582	T21978	T21979	mark	whereas,were
R6583	T21979	T21969	advcl	were,amplify
R6584	T21980	T21981	det	the,primers
R6585	T21981	T21979	nsubj	primers,were
R6586	T21982	T21981	compound	antisense,primers
R6587	T21983	T21984	npadvmod	allele,specific
R6588	T21984	T21979	acomp	specific,were
R6589	T21985	T21984	punct	-,specific
R659	T3633	T3634	prep	To,suggests
R6590	T21986	T21966	punct	.,used
R6591	T21988	T21989	det	The,primers
R6592	T21989	T21990	nsubjpass	primers,depicted
R6593	T21991	T21989	punct	", ",primers
R6594	T21992	T21989	acl	represented,primers
R6595	T21993	T21992	agent	by,represented
R6596	T21994	T21993	pobj	triangles,by
R6597	T21995	T21990	punct	", ",depicted
R6598	T21996	T21990	auxpass	are,depicted
R6599	T21997	T21990	prep	in,depicted
R66	T738	T736	amod	regulatory,subunit
R660	T3642	T3640	prep	between,interplay
R6600	T21998	T21999	nummod	two,sets
R6601	T21999	T21997	pobj	sets,in
R6602	T22000	T21999	punct	(,sets
R6603	T22001	T21999	amod	nested,sets
R6604	T22002	T21999	punct	),sets
R6605	T22003	T21999	prep	for,sets
R6606	T22004	T22003	pobj	each,for
R6607	T22005	T22004	prep	of,each
R6608	T22006	T22007	det	the,reactions
R6609	T22007	T22005	pobj	reactions,of
R661	T3635	T3633	pobj	date,To
R6610	T22008	T22007	nummod	three,reactions
R6611	T22009	T21990	punct	.,depicted
R6612	T22011	T22012	nsubjpass	Exons,represented
R6613	T22012	T22014	ccomp	represented,cassette
R6614	T22013	T22012	auxpass	are,represented
R6615	T22015	T22012	agent	by,represented
R6616	T22016	T22017	amod	open,rectangles
R6617	T22017	T22015	pobj	rectangles,by
R6618	T22018	T22017	punct	", ",rectangles
R6619	T22019	T22017	amod	vertical,rectangles
R662	T3636	T3634	punct	", ",suggests
R6620	T22020	T22014	punct	", ",cassette
R6621	T22021	T22014	det	the,cassette
R6622	T22022	T22023	compound	PGK,Neo
R6623	T22023	T22014	compound	Neo,cassette
R6624	T22024	T22023	punct	-,Neo
R6625	T22025	T22014	agent	by,cassette
R6626	T22026	T22027	det	an,rectangle
R6627	T22027	T22025	pobj	rectangle,by
R6628	T22028	T22027	amod	open,rectangle
R6629	T22029	T22027	punct	", ",rectangle
R663	T3643	T3642	pobj	caspases,between
R6630	T22030	T22027	amod	horizontal,rectangle
R6631	T22031	T22014	punct	", ",cassette
R6632	T22032	T22033	mark	while,denote
R6633	T22033	T22014	advcl	denote,cassette
R6634	T22034	T22035	amod	thin,lines
R6635	T22035	T22033	nsubj	lines,denote
R6636	T22036	T22035	amod	vertical,lines
R6637	T22037	T22038	det	the,boundaries
R6638	T22038	T22033	dobj	boundaries,denote
R6639	T22039	T22038	prep	of,boundaries
R664	T3637	T3638	det	the,literature
R6640	T22040	T22041	det	the,arm
R6641	T22041	T22039	pobj	arm,of
R6642	T22042	T22041	amod	short,arm
R6643	T22043	T22038	cc	and,boundaries
R6644	T22044	T22045	det	the,end
R6645	T22045	T22038	conj	end,boundaries
R6646	T22046	T22045	nummod	5,end
R6647	T22047	T22046	punct	',5
R6648	T22048	T22045	prep	of,end
R6649	T22049	T22050	det	the,arm
R665	T3638	T3634	nsubj	literature,suggests
R6650	T22050	T22048	pobj	arm,of
R6651	T22051	T22050	amod	long,arm
R6652	T22052	T22050	punct	(,arm
R6653	T22053	T22050	amod	downstream,arm
R6654	T22054	T22050	punct	),arm
R6655	T22055	T22014	punct	.,cassette
R6656	T22057	T22058	det	A,rectangle
R6657	T22058	T22062	nsubj	rectangle,delineates
R6658	T22059	T22058	amod	grey,rectangle
R6659	T22060	T22058	punct	", ",rectangle
R666	T3639	T3640	det	a,interplay
R6660	T22061	T22058	amod	horizontal,rectangle
R6661	T22063	T22064	det	the,segment
R6662	T22064	T22062	dobj	segment,delineates
R6663	T22065	T22064	prep	of,segment
R6664	T22066	T22067	det	the,allele
R6665	T22067	T22065	pobj	allele,of
R6666	T22068	T22069	amod	wild,type
R6667	T22069	T22067	compound	type,allele
R6668	T22070	T22069	punct	-,type
R6669	T22071	T22072	dep	that,replaced
R667	T3644	T3643	cc	and,caspases
R6670	T22072	T22064	relcl	replaced,segment
R6671	T22073	T22072	auxpass	is,replaced
R6672	T22074	T22072	agent	by,replaced
R6673	T22075	T22076	det	the,cassette
R6674	T22076	T22074	pobj	cassette,by
R6675	T22077	T22078	compound	PGK,Neo
R6676	T22078	T22076	compound	Neo,cassette
R6677	T22079	T22078	punct	-,Neo
R6678	T22080	T22072	prep	in,replaced
R6679	T22081	T22082	det	the,allele
R668	T3645	T3643	conj	calpains,caspases
R6680	T22082	T22080	pobj	allele,in
R6681	T22083	T22082	compound	mutant,allele
R6682	T22084	T22062	punct	.,delineates
R6683	T22641	T22640	prep	of,Genotyping
R6684	T22642	T22643	amod	pre-implantation,embryos
R6685	T22643	T22641	pobj	embryos,of
R6686	T22644	T22640	punct	.,Genotyping
R6687	T22646	T22647	det	A,strategy
R6688	T22647	T22650	nsubjpass	strategy,used
R6689	T22648	T22647	amod	nested,strategy
R669	T3646	T3647	punct	[,24
R6690	T22649	T22647	compound	PCR,strategy
R6691	T22651	T22650	auxpass	was,used
R6692	T22652	T22653	aux	to,genotype
R6693	T22653	T22650	advcl	genotype,used
R6694	T22654	T22653	dobj	embryos,genotype
R6695	T22655	T22656	amod	prior,to
R6696	T22656	T22653	prep	to,genotype
R6697	T22657	T22656	pobj	implantation,to
R6698	T22658	T22659	mark	as,described
R6699	T22659	T22653	advcl	described,genotype
R67	T739	T736	acl	encoded,subunit
R670	T3647	T3640	parataxis	24,interplay
R6700	T22660	T22659	prep	in,described
R6701	T22661	T22660	pobj	Figure,in
R6702	T22662	T22661	nummod	4,Figure
R6703	T22663	T22650	punct	.,used
R6704	T22665	T22666	nmod	Capn2,mice
R6705	T22666	T22670	nsubjpass	mice,mated
R6706	T22667	T22665	punct	+,Capn2
R6707	T22668	T22665	punct	/,Capn2
R6708	T22669	T22665	punct	-,Capn2
R6709	T22671	T22670	auxpass	were,mated
R671	T3746	T3745	punct	-,calpain
R6710	T22672	T22670	cc	and,mated
R6711	T22673	T22674	det	the,date
R6712	T22674	T22675	nsubj	date,established
R6713	T22675	T22670	conj	established,mated
R6714	T22676	T22674	prep	of,date
R6715	T22677	T22676	pobj	fertilization,of
R6716	T22678	T22675	agent	by,established
R6717	T22679	T22680	det	the,appearance
R6718	T22680	T22678	pobj	appearance,by
R6719	T22681	T22680	prep	of,appearance
R672	T3747	T3742	prep	in,role
R6720	T22682	T22683	amod	vaginal,plugs
R6721	T22683	T22681	pobj	plugs,of
R6722	T22684	T22675	punct	.,established
R6723	T22686	T22687	nmod	Blastocyst,embryos
R6724	T22687	T22695	nsubjpass	embryos,flushed
R6725	T22688	T22689	punct	(,E3.5
R6726	T22689	T22686	parataxis	E3.5,Blastocyst
R6727	T22690	T22689	punct	),E3.5
R6728	T22691	T22686	cc	or,Blastocyst
R6729	T22692	T22693	nummod	8,cell
R673	T3648	T3647	nummod	23,24
R6730	T22693	T22686	conj	cell,Blastocyst
R6731	T22694	T22693	punct	-,cell
R6732	T22696	T22697	punct	(,E2.5
R6733	T22697	T22687	parataxis	E2.5,embryos
R6734	T22698	T22697	punct	),E2.5
R6735	T22699	T22695	auxpass	were,flushed
R6736	T22700	T22695	prep	from,flushed
R6737	T22701	T22702	det	the,oviducts
R6738	T22702	T22700	pobj	oviducts,from
R6739	T22703	T22695	cc	and,flushed
R674	T3748	T3749	amod	epidermal,factor
R6740	T22704	T22705	advmod	then,digested
R6741	T22705	T22695	conj	digested,flushed
R6742	T22706	T22705	prep	with,digested
R6743	T22707	T22708	compound	proteinase,K
R6744	T22708	T22706	pobj	K,with
R6745	T22709	T22695	punct	.,flushed
R6746	T22711	T22712	prep	In,co-amplified
R6747	T22713	T22714	amod	separate,segments
R6748	T22714	T22711	pobj	segments,In
R6749	T22715	T22714	compound	reactions,segments
R675	T3749	T3751	npadvmod	factor,induced
R6750	T22716	T22714	acl	found,segments
R6751	T22717	T22716	advmod	exclusively,found
R6752	T22718	T22716	prep	in,found
R6753	T22719	T22720	preconj	either,type
R6754	T22720	T22718	pobj	type,in
R6755	T22721	T22720	det	the,type
R6756	T22722	T22720	amod	wild,type
R6757	T22723	T22720	punct	-,type
R6758	T22724	T22720	cc	or,type
R6759	T22725	T22720	conj	mutant,type
R676	T3750	T3749	compound	growth,factor
R6760	T22726	T22712	nsubjpass	alleles,co-amplified
R6761	T22727	T22712	auxpass	were,co-amplified
R6762	T22728	T22712	prep	with,co-amplified
R6763	T22729	T22730	det	an,sequence
R6764	T22730	T22728	pobj	sequence,with
R6765	T22731	T22730	amod	internal,sequence
R6766	T22732	T22730	compound	control,sequence
R6767	T22733	T22730	punct	", ",sequence
R6768	T22734	T22730	acl	located,sequence
R6769	T22735	T22734	prep	in,located
R677	T3649	T3647	punct	",",24
R6770	T22736	T22737	det	the,arm
R6771	T22737	T22735	pobj	arm,in
R6772	T22738	T22737	amod	short,arm
R6773	T22739	T22737	punct	(,arm
R6774	T22740	T22737	amod	upstream,arm
R6775	T22741	T22737	punct	),arm
R6776	T22742	T22737	prep	of,arm
R6777	T22743	T22744	det	the,vector
R6778	T22744	T22742	pobj	vector,of
R6779	T22745	T22744	compound	targeting,vector
R678	T3751	T3756	amod	induced,motility
R6780	T22746	T22744	punct	", ",vector
R6781	T22747	T22748	dep	which,found
R6782	T22748	T22744	relcl	found,vector
R6783	T22749	T22748	auxpass	is,found
R6784	T22750	T22748	prep	in,found
R6785	T22751	T22752	det	both,alleles
R6786	T22752	T22750	pobj	alleles,in
R6787	T22753	T22712	punct	.,co-amplified
R6788	T22755	T22756	det	The,products
R6789	T22756	T22758	nsubj	products,were
R679	T3752	T3749	punct	(,factor
R6790	T22757	T22756	amod	final,products
R6791	T22759	T22760	nummod	429,bp
R6792	T22760	T22758	attr	bp,were
R6793	T22761	T22758	prep	for,were
R6794	T22762	T22763	det	the,allele
R6795	T22763	T22761	pobj	allele,for
R6796	T22764	T22765	amod	wild,type
R6797	T22765	T22763	compound	type,allele
R6798	T22766	T22765	punct	-,type
R6799	T22767	T22758	punct	", ",were
R68	T740	T739	agent	by,encoded
R680	T3650	T3647	punct	],24
R6800	T22768	T22769	nummod	389,bp
R6801	T22769	T22758	conj	bp,were
R6802	T22770	T22769	prep	for,bp
R6803	T22771	T22772	det	the,allele
R6804	T22772	T22770	pobj	allele,for
R6805	T22773	T22772	compound	mutant,allele
R6806	T22774	T22769	punct	", ",bp
R6807	T22775	T22769	cc	and,bp
R6808	T22776	T22777	nummod	213,bp
R6809	T22777	T22769	conj	bp,bp
R681	T3753	T3749	appos	EGF,factor
R6810	T22778	T22777	prep	for,bp
R6811	T22779	T22780	det	the,control
R6812	T22780	T22778	pobj	control,for
R6813	T22781	T22780	amod	internal,control
R6814	T22782	T22758	punct	.,were
R6815	T22784	T22785	punct	(,A
R6816	T22785	T22786	meta	A,example
R6817	T22787	T22785	punct	),A
R6818	T22788	T22786	det	A,example
R6819	T22789	T22786	amod	representative,example
R682	T3754	T3751	punct	),induced
R6820	T22790	T22786	prep	of,example
R6821	T22791	T22792	det	the,genotyping
R6822	T22792	T22790	pobj	genotyping,of
R6823	T22793	T22792	prep	of,genotyping
R6824	T22794	T22795	compound	blastocyst,embryos
R6825	T22795	T22793	pobj	embryos,of
R6826	T22796	T22795	compound	stage,embryos
R6827	T22797	T22786	punct	.,example
R6828	T22799	T22800	nmod	Embryos,1
R6829	T22800	T22802	nsubj	1,were
R683	T3755	T3751	punct	-,induced
R6830	T22801	T22800	punct	#,1
R6831	T22803	T22800	punct	", ",1
R6832	T22804	T22800	conj	2,1
R6833	T22805	T22804	punct	", ",2
R6834	T22806	T22804	conj	4,2
R6835	T22807	T22806	punct	", ",4
R6836	T22808	T22806	conj	5,4
R6837	T22809	T22808	punct	", ",5
R6838	T22810	T22808	cc	and,5
R6839	T22811	T22808	conj	6,5
R684	T3651	T3640	cc	and,interplay
R6840	T22812	T22802	attr	Capn2,were
R6841	T22813	T22812	punct	+,Capn2
R6842	T22814	T22812	punct	/,Capn2
R6843	T22815	T22812	punct	-,Capn2
R6844	T22816	T22817	mark	whereas,were
R6845	T22817	T22802	advcl	were,were
R6846	T22818	T22819	nmod	embryos,3
R6847	T22819	T22817	nsubj	3,were
R6848	T22820	T22819	punct	#,3
R6849	T22821	T22819	cc	and,3
R685	T3756	T3747	pobj	motility,in
R6850	T22822	T22823	punct	#,7
R6851	T22823	T22819	conj	7,3
R6852	T22824	T22817	attr	Capn2,were
R6853	T22825	T22824	punct	+,Capn2
R6854	T22826	T22824	punct	/,Capn2
R6855	T22827	T22824	punct	+,Capn2
R6856	T22828	T22817	punct	", ",were
R6857	T22829	T22817	advcl	denoted,were
R6858	T22830	T22829	agent	by,denoted
R6859	T22831	T22832	det	the,absence
R686	T3757	T3756	compound	cell,motility
R6860	T22832	T22830	pobj	absence,by
R6861	T22833	T22832	prep	of,absence
R6862	T22834	T22835	det	the,signal
R6863	T22835	T22833	pobj	signal,of
R6864	T22836	T22837	nummod	389,bp
R6865	T22837	T22835	compound	bp,signal
R6866	T22838	T22835	compound	mutant,signal
R6867	T22839	T22802	punct	.,were
R6868	T22841	T22842	punct	(,B
R6869	T22842	T22843	meta	B,shown
R687	T3758	T3759	punct	[,29
R6870	T22844	T22842	punct	),B
R6871	T22845	T22846	det	An,example
R6872	T22846	T22843	nsubjpass	example,shown
R6873	T22847	T22846	prep	of,example
R6874	T22848	T22849	det	the,genotyping
R6875	T22849	T22847	pobj	genotyping,of
R6876	T22850	T22849	prep	of,genotyping
R6877	T22851	T22852	nummod	8,cell
R6878	T22852	T22854	compound	cell,embryos
R6879	T22853	T22852	punct	-,cell
R688	T3759	T3742	parataxis	29,role
R6880	T22854	T22850	pobj	embryos,of
R6881	T22855	T22843	auxpass	is,shown
R6882	T22856	T22843	punct	.,shown
R6883	T22858	T22859	nmod	Embryos,1
R6884	T22859	T22861	nsubj	1,were
R6885	T22860	T22859	punct	#,1
R6886	T22862	T22859	punct	", ",1
R6887	T22863	T22859	conj	2,1
R6888	T22864	T22863	punct	", ",2
R6889	T22865	T22863	conj	3,2
R689	T3760	T3759	nummod	28,29
R6890	T22866	T22865	punct	", ",3
R6891	T22867	T22865	conj	5,3
R6892	T22868	T22867	punct	", ",5
R6893	T22869	T22867	cc	and,5
R6894	T22870	T22867	conj	6,5
R6895	T22871	T22861	attr	Capn2,were
R6896	T22872	T22871	punct	+,Capn2
R6897	T22873	T22871	punct	/,Capn2
R6898	T22874	T22871	punct	-,Capn2
R6899	T22875	T22876	mark	while,was
R69	T741	T740	pobj	Capn4,by
R690	T3761	T3759	punct	",",29
R6900	T22876	T22861	advcl	was,were
R6901	T22877	T22876	nsubj	embryo,was
R6902	T22878	T22877	punct	#,embryo
R6903	T22879	T22877	nummod	4,embryo
R6904	T22880	T22876	attr	Capn2,was
R6905	T22881	T22880	punct	-,Capn2
R6906	T22882	T22880	punct	/,Capn2
R6907	T22883	T22880	punct	-,Capn2
R6908	T22884	T22880	punct	", ",Capn2
R6909	T22885	T22880	acl	marked,Capn2
R691	T3652	T3640	conj	impact,interplay
R6910	T22886	T22885	agent	by,marked
R6911	T22887	T22888	det	the,absence
R6912	T22888	T22886	pobj	absence,by
R6913	T22889	T22888	prep	of,absence
R6914	T22890	T22891	det	the,signal
R6915	T22891	T22889	pobj	signal,of
R6916	T22892	T22893	nummod	429,bp
R6917	T22893	T22891	nmod	bp,signal
R6918	T22894	T22895	amod	wild,type
R6919	T22895	T22891	compound	type,signal
R692	T3762	T3759	punct	],29
R6920	T22896	T22895	punct	-,type
R6921	T22897	T22861	punct	.,were
R6922	T22899	T22900	punct	(,M
R6923	T22900	T22901	nsubj	M,denotes
R6924	T22902	T22900	punct	),M
R6925	T22903	T22904	det	the,marker
R6926	T22904	T22901	dobj	marker,denotes
R6927	T22905	T22906	amod	molecular,weight
R6928	T22906	T22904	compound	weight,marker
R6929	T22907	T22901	punct	.,denotes
R693	T3763	T3742	cc	and,role
R6930	T22936	T22937	compound	Genotype,distribution
R6931	T22938	T22937	prep	of,distribution
R6932	T22939	T22938	pobj	offspring,of
R6933	T22940	T22939	acl	derived,offspring
R6934	T22941	T22940	prep	from,derived
R6935	T22942	T22943	nmod	Capn2,mice
R6936	T22943	T22941	pobj	mice,from
R6937	T22944	T22943	amod	transgenic,mice
R6938	T22945	T22937	punct	.,distribution
R6939	T23423	T23424	compound	Targeting,strategy
R694	T3764	T3765	det	a,role
R6940	T23425	T23424	prep	for,strategy
R6941	T23426	T23425	pobj	disruption,for
R6942	T23427	T23426	prep	of,disruption
R6943	T23428	T23429	det	the,gene
R6944	T23429	T23427	pobj	gene,of
R6945	T23430	T23429	amod	murine,gene
R6946	T23431	T23429	compound	Capn2,gene
R6947	T23432	T23424	punct	.,strategy
R6948	T23434	T23435	det	The,gene
R6949	T23435	T23438	nsubjpass	gene,disrupted
R695	T3653	T3652	prep	of,impact
R6950	T23436	T23435	amod	murine,gene
R6951	T23437	T23435	compound	Capn2,gene
R6952	T23439	T23435	punct	", ",gene
R6953	T23440	T23435	acl	encoding,gene
R6954	T23441	T23442	det	the,subunit
R6955	T23442	T23440	dobj	subunit,encoding
R6956	T23443	T23444	nmod	m,k
R6957	T23444	T23442	compound	k,subunit
R6958	T23445	T23444	punct	-,k
R6959	T23446	T23444	nummod	80,k
R696	T3765	T3742	conj	role,role
R6960	T23447	T23438	punct	", ",disrupted
R6961	T23448	T23438	auxpass	was,disrupted
R6962	T23449	T23438	prep	in,disrupted
R6963	T23450	T23451	compound	ES,cells
R6964	T23451	T23449	pobj	cells,in
R6965	T23452	T23438	agent	by,disrupted
R6966	T23453	T23454	amod	homologous,recombination
R6967	T23454	T23452	pobj	recombination,by
R6968	T23455	T23438	punct	.,disrupted
R6969	T23457	T23458	det	The,structures
R697	T3766	T3765	prep	for,role
R6970	T23458	T23459	nsubjpass	structures,depicted
R6971	T23460	T23458	prep	of,structures
R6972	T23461	T23462	det	the,allele
R6973	T23462	T23460	pobj	allele,of
R6974	T23463	T23464	amod	wild,type
R6975	T23464	T23462	compound	type,allele
R6976	T23465	T23464	punct	-,type
R6977	T23466	T23467	punct	(,top
R6978	T23467	T23462	parataxis	top,allele
R6979	T23468	T23467	punct	),top
R698	T3654	T3653	pobj	calpain,of
R6980	T23469	T23462	punct	", ",allele
R6981	T23470	T23471	det	the,vector
R6982	T23471	T23462	conj	vector,allele
R6983	T23472	T23471	compound	targeting,vector
R6984	T23473	T23474	punct	(,middle
R6985	T23474	T23471	parataxis	middle,vector
R6986	T23475	T23474	punct	),middle
R6987	T23476	T23471	punct	", ",vector
R6988	T23477	T23471	cc	and,vector
R6989	T23478	T23479	det	the,allele
R699	T3767	T3768	compound	μ,calpain
R6990	T23479	T23471	conj	allele,vector
R6991	T23480	T23479	compound	mutant,allele
R6992	T23481	T23482	punct	(,bottom
R6993	T23482	T23479	parataxis	bottom,allele
R6994	T23483	T23482	punct	),bottom
R6995	T23484	T23459	auxpass	are,depicted
R6996	T23485	T23459	punct	.,depicted
R6997	T23487	T23488	prep	In,replaces
R6998	T23488	T23498	ccomp	replaces,flanked
R6999	T23489	T23490	det	the,allele
R7	T671	T669	prep	for,required
R70	T742	T711	punct	.,composed
R700	T3768	T3766	pobj	calpain,for
R7000	T23490	T23487	pobj	allele,In
R7001	T23491	T23490	compound	mutant,allele
R7002	T23492	T23488	punct	", ",replaces
R7003	T23493	T23494	det	a,cassette
R7004	T23494	T23488	nsubj	cassette,replaces
R7005	T23495	T23496	compound	PGK,Neo
R7006	T23496	T23494	compound	Neo,cassette
R7007	T23497	T23496	punct	-,Neo
R7008	T23499	T23500	det	a,fragment
R7009	T23500	T23488	dobj	fragment,replaces
R701	T3769	T3768	punct	-,calpain
R7010	T23501	T23502	nummod	0.8,kb
R7011	T23502	T23500	nmod	kb,fragment
R7012	T23503	T23502	punct	-,kb
R7013	T23504	T23500	amod	genomic,fragment
R7014	T23505	T23500	acl	containing,fragment
R7015	T23506	T23505	dobj	exon,containing
R7016	T23507	T23506	nummod	7,exon
R7017	T23508	T23509	punct	(,rectangle
R7018	T23509	T23506	parataxis	rectangle,exon
R7019	T23510	T23509	amod	grey,rectangle
R702	T3655	T3652	prep	on,impact
R7020	T23511	T23509	punct	),rectangle
R7021	T23512	T23513	dep	which,encodes
R7022	T23513	T23500	relcl	encodes,fragment
R7023	T23514	T23515	det	the,residue
R7024	T23515	T23513	dobj	residue,encodes
R7025	T23516	T23515	amod	active,residue
R7026	T23517	T23515	compound	site,residue
R7027	T23518	T23515	compound	asparagine,residue
R7028	T23519	T23515	punct	(,residue
R7029	T23520	T23515	appos	Asn286,residue
R703	T3770	T3765	prep	in,role
R7030	T23521	T23498	punct	),flanked
R7031	T23522	T23498	prep	In,flanked
R7032	T23523	T23524	det	the,vector
R7033	T23524	T23522	pobj	vector,In
R7034	T23525	T23524	compound	targeting,vector
R7035	T23526	T23498	punct	", ",flanked
R7036	T23527	T23528	det	the,cassette
R7037	T23528	T23498	nsubjpass	cassette,flanked
R7038	T23529	T23530	compound	PGK,Neo
R7039	T23530	T23528	compound	Neo,cassette
R704	T3771	T3772	npadvmod	interferon,inducible
R7040	T23531	T23530	punct	-,Neo
R7041	T23532	T23498	auxpass	is,flanked
R7042	T23533	T23498	agent	by,flanked
R7043	T23534	T23535	nummod	2.7,kb
R7044	T23535	T23533	pobj	kb,by
R7045	T23536	T23535	punct	-,kb
R7046	T23537	T23535	prep	of,kb
R7047	T23538	T23539	nmod	Capn2,sequence
R7048	T23539	T23537	pobj	sequence,of
R7049	T23540	T23539	amod	homologous,sequence
R705	T3772	T3774	amod	inducible,migration
R7050	T23541	T23498	prep	in,flanked
R7051	T23542	T23543	det	the,arm
R7052	T23543	T23541	pobj	arm,in
R7053	T23544	T23543	amod	upstream,arm
R7054	T23545	T23543	punct	(,arm
R7055	T23546	T23543	amod	short,arm
R7056	T23547	T23543	punct	),arm
R7057	T23548	T23498	cc	and,flanked
R7058	T23549	T23550	nummod	7.9,kb
R7059	T23550	T23498	conj	kb,flanked
R706	T3656	T3657	compound	cell,death
R7060	T23551	T23550	punct	-,kb
R7061	T23552	T23550	prep	of,kb
R7062	T23553	T23552	pobj	homology,of
R7063	T23554	T23550	prep	in,kb
R7064	T23555	T23556	det	the,arm
R7065	T23556	T23554	pobj	arm,in
R7066	T23557	T23556	amod	downstream,arm
R7067	T23558	T23556	punct	(,arm
R7068	T23559	T23556	amod	long,arm
R7069	T23560	T23556	punct	),arm
R707	T3773	T3772	punct	-,inducible
R7070	T23561	T23498	punct	.,flanked
R7071	T23563	T23564	det	A,probe
R7072	T23564	T23565	nsubj	probe,detects
R7073	T23566	T23564	acl	located,probe
R7074	T23567	T23568	advmod	immediately,outside
R7075	T23568	T23566	advmod	outside,located
R7076	T23569	T23568	prep	of,outside
R7077	T23570	T23571	det	the,arm
R7078	T23571	T23569	pobj	arm,of
R7079	T23572	T23571	amod	short,arm
R708	T3774	T3770	pobj	migration,in
R7080	T23573	T23574	det	a,fragment
R7081	T23574	T23565	dobj	fragment,detects
R7082	T23575	T23576	nummod	3.5,kb
R7083	T23576	T23574	compound	kb,fragment
R7084	T23577	T23576	punct	-,kb
R7085	T23578	T23574	compound	BamHI,fragment
R7086	T23579	T23565	prep	from,detects
R7087	T23580	T23581	det	the,allele
R7088	T23581	T23579	pobj	allele,from
R7089	T23582	T23583	amod	wild,type
R709	T3775	T3776	npadvmod	protein,induced
R7090	T23583	T23581	compound	type,allele
R7091	T23584	T23583	punct	-,type
R7092	T23585	T23565	cc	and,detects
R7093	T23586	T23587	det	a,fragment
R7094	T23587	T23565	conj	fragment,detects
R7095	T23588	T23589	nummod	5.3,kb
R7096	T23589	T23587	compound	kb,fragment
R7097	T23590	T23589	punct	-,kb
R7098	T23591	T23587	compound	BamHI,fragment
R7099	T23592	T23587	prep	from,fragment
R71	T744	T745	nsubj	Disruption,abolished
R710	T3657	T3658	compound	death,components
R7100	T23593	T23594	det	the,allele
R7101	T23594	T23592	pobj	allele,from
R7102	T23595	T23594	compound	mutant,allele
R7103	T23596	T23565	punct	.,detects
R7104	T23598	T23599	nsubjpass	Exons,depicted
R7105	T23600	T23599	auxpass	are,depicted
R7106	T23601	T23599	prep	as,depicted
R7107	T23602	T23603	amod	open,rectangles
R7108	T23603	T23601	pobj	rectangles,as
R7109	T23604	T23603	amod	vertical,rectangles
R711	T3776	T3774	amod	induced,migration
R7110	T23605	T23599	prep	except,depicted
R7111	T23606	T23605	prep	for,except
R7112	T23607	T23606	pobj	exon,for
R7113	T23608	T23607	nummod	7,exon
R7114	T23609	T23610	dep	which,represented
R7115	T23610	T23607	relcl	represented,exon
R7116	T23611	T23610	auxpass	is,represented
R7117	T23612	T23610	agent	by,represented
R7118	T23613	T23614	det	a,rectangle
R7119	T23614	T23612	pobj	rectangle,by
R712	T3777	T3775	nummod	9,protein
R7120	T23615	T23614	amod	solid,rectangle
R7121	T23616	T23614	amod	vertical,rectangle
R7122	T23617	T23599	punct	.,depicted
R7123	T23619	T23620	det	The,probe
R7124	T23620	T23621	nsubjpass	probe,shown
R7125	T23622	T23620	acl	used,probe
R7126	T23623	T23622	prep	in,used
R7127	T23624	T23625	amod	most,analyses
R7128	T23625	T23623	pobj	analyses,in
R7129	T23626	T23627	compound	Southern,blot
R713	T3778	T3776	punct	-,induced
R7130	T23627	T23625	compound	blot,analyses
R7131	T23628	T23621	auxpass	is,shown
R7132	T23629	T23621	prep	as,shown
R7133	T23630	T23631	det	a,rectangle
R7134	T23631	T23629	pobj	rectangle,as
R7135	T23632	T23631	amod	solid,rectangle
R7136	T23633	T23631	punct	", ",rectangle
R7137	T23634	T23631	amod	horizontal,rectangle
R7138	T23635	T23621	punct	", ",shown
R7139	T23636	T23637	mark	while,mark
R714	T3658	T3655	pobj	components,on
R7140	T23637	T23621	advcl	mark,shown
R7141	T23638	T23637	nsubj	triangles,mark
R7142	T23639	T23640	det	the,positions
R7143	T23640	T23637	dobj	positions,mark
R7144	T23641	T23640	prep	of,positions
R7145	T23642	T23643	compound	PCR,primers
R7146	T23643	T23641	pobj	primers,of
R7147	T23644	T23645	advmod	also,used
R7148	T23645	T23643	acl	used,primers
R7149	T23646	T23645	prep	for,used
R715	T3779	T3774	prep	of,migration
R7150	T23647	T23648	compound	genotyping,purposes
R7151	T23648	T23646	pobj	purposes,for
R7152	T23649	T23621	punct	.,shown
R7153	T23673	T23674	compound	Oligonucleotide,primers
R7154	T23675	T23674	acl	used,primers
R7155	T23676	T23677	aux	to,genotype
R7156	T23677	T23675	advcl	genotype,used
R7157	T23678	T23679	det	the,locus
R7158	T23679	T23677	dobj	locus,genotype
R7159	T23680	T23679	compound	Capn2,locus
R716	T3780	T3779	pobj	keratinocytes,of
R717	T3781	T3782	punct	[,28
R718	T3659	T3658	compound	pathway,components
R719	T3782	T3765	parataxis	28,role
R72	T746	T744	prep	of,Disruption
R720	T3783	T3782	punct	],28
R721	T3784	T3733	punct	.,suggested
R722	T3660	T3661	punct	[,25
R723	T3786	T3787	det	A,inhibitor
R724	T3661	T3652	parataxis	25,impact
R725	T3787	T3791	nsubjpass	inhibitor,used
R726	T3788	T3789	npadvmod	cell,permeable
R727	T3789	T3787	amod	permeable,inhibitor
R728	T3790	T3787	compound	calpain,inhibitor
R729	T3662	T3661	punct	],25
R73	T747	T748	det	the,gene
R730	T3792	T3787	punct	(,inhibitor
R731	T3663	T3634	punct	.,suggests
R732	T3793	T3794	dep	which,inhibits
R733	T3794	T3787	relcl	inhibits,inhibitor
R734	T3795	T3794	advmod	likely,inhibits
R735	T3665	T3666	det	The,lack
R736	T3796	T3797	amod	other,proteases
R737	T3797	T3794	dobj	proteases,inhibits
R738	T3798	T3797	compound	thiol,proteases
R739	T3799	T3797	punct	-,proteases
R74	T748	T746	pobj	gene,of
R740	T3800	T3801	advmod	as,well
R741	T3801	T3794	advmod	well,inhibits
R742	T3802	T3791	punct	),used
R743	T3666	T3667	nsubj	lack,contributes
R744	T3803	T3791	aux	has,used
R745	T3804	T3791	auxpass	been,used
R746	T3805	T3806	aux	to,select
R747	T3668	T3666	prep	of,lack
R748	T3806	T3791	advcl	select,used
R749	T3807	T3806	dobj	cells,select
R75	T749	T748	compound	mouse,gene
R750	T3808	T3807	acl	lacking,cells
R751	T3809	T3810	compound	μ,calpain
R752	T3669	T3670	advmod	highly,specific
R753	T3670	T3671	amod	specific,inhibitors
R754	T3810	T3808	dobj	calpain,lacking
R755	T3671	T3668	pobj	inhibitors,of
R756	T3811	T3810	punct	-,calpain
R757	T3812	T3813	dep	which,display
R758	T3813	T3807	relcl	display,cells
R759	T3672	T3673	npadvmod	cell,permeable
R76	T750	T748	compound	Capn4,gene
R760	T3814	T3815	amod	reduced,rates
R761	T3815	T3813	dobj	rates,display
R762	T3816	T3815	compound	proliferation,rates
R763	T3673	T3671	amod	permeable,inhibitors
R764	T3817	T3818	punct	[,30
R765	T3818	T3813	parataxis	30,display
R766	T3819	T3818	punct	],30
R767	T3674	T3673	punct	-,permeable
R768	T3820	T3791	punct	.,used
R769	T3675	T3671	prep	of,inhibitors
R77	T751	T752	preconj	both,calpain
R770	T3822	T3823	advmod	Interestingly,persisted
R771	T3676	T3675	pobj	calpains,of
R772	T3824	T3823	punct	", ",persisted
R773	T3825	T3826	compound	m,calpain
R774	T3826	T3828	compound	calpain,expression
R775	T3677	T3667	prep	to,contributes
R776	T3827	T3826	punct	-,calpain
R777	T3828	T3823	nsubj	expression,persisted
R778	T3829	T3823	prep	in,persisted
R779	T3678	T3679	det	the,challenge
R78	T752	T755	nmod	calpain,activity
R780	T3830	T3831	det	these,cells
R781	T3831	T3829	pobj	cells,in
R782	T3832	T3823	punct	", ",persisted
R783	T3679	T3677	pobj	challenge,to
R784	T3833	T3823	advcl	suggesting,persisted
R785	T3834	T3835	det	a,requirement
R786	T3835	T3833	dobj	requirement,suggesting
R787	T3680	T3679	prep	of,challenge
R788	T3836	T3835	amod	possible,requirement
R789	T3837	T3835	prep	of,requirement
R79	T753	T752	nmod	μ,calpain
R790	T3681	T3680	pcomp	investigating,of
R791	T3838	T3839	compound	m,calpain
R792	T3839	T3837	pobj	calpain,of
R793	T3840	T3839	punct	-,calpain
R794	T3841	T3835	prep	for,requirement
R795	T3842	T3843	compound	cell,survival
R796	T3843	T3841	pobj	survival,for
R797	T3682	T3681	cc	and,investigating
R798	T3844	T3845	punct	[,30
R799	T3845	T3823	parataxis	30,persisted
R8	T672	T673	nmod	preimplantation,development
R80	T754	T752	punct	-,calpain
R800	T3846	T3845	punct	],30
R801	T3683	T3681	conj	defining,investigating
R802	T3847	T3823	punct	.,persisted
R803	T3684	T3685	compound	calpain,functions
R804	T3849	T3850	amod	Targeted,deletion
R805	T3685	T3683	dobj	functions,defining
R806	T3686	T3683	prep	in,defining
R807	T3850	T3852	nsubj	deletion,provides
R808	T3687	T3688	det	these,processes
R809	T3851	T3850	compound	gene,deletion
R81	T755	T745	dobj	activity,abolished
R810	T3853	T3850	prep	in,deletion
R811	T3688	T3686	pobj	processes,in
R812	T3854	T3853	pobj	mice,in
R813	T3855	T3856	det	a,approach
R814	T3689	T3667	punct	.,contributes
R815	T3856	T3852	dobj	approach,provides
R816	T3691	T3692	mark	Although,provides
R817	T3857	T3856	amod	powerful,approach
R818	T3858	T3856	prep	to,approach
R819	T3859	T3858	pcomp	determining,to
R82	T756	T752	cc	and,calpain
R820	T3860	T3861	det	the,roles
R821	T3692	T3708	advcl	provides,distinguish
R822	T3861	T3859	dobj	roles,determining
R823	T3862	T3861	amod	physiological,roles
R824	T3863	T3861	prep	of,roles
R825	T3693	T3692	nsubj	over-expression,provides
R826	T3864	T3865	nmod	μ,calpain
R827	T3865	T3863	pobj	calpain,of
R828	T3694	T3693	prep	of,over-expression
R829	T3866	T3864	punct	-,μ
R83	T757	T758	compound	m,calpain
R830	T3867	T3864	cc	and,μ
R831	T3868	T3864	conj	m,μ
R832	T3695	T3694	pobj	calpastatin,of
R833	T3869	T3865	punct	-,calpain
R834	T3870	T3856	cc	and,approach
R835	T3871	T3872	det	the,opportunity
R836	T3696	T3695	punct	", ",calpastatin
R837	T3872	T3856	conj	opportunity,approach
R838	T3873	T3874	aux	to,approach
R839	T3697	T3698	det	the,inhibitor
R84	T758	T752	conj	calpain,calpain
R840	T3874	T3872	acl	approach,opportunity
R841	T3875	T3876	poss	their,functions
R842	T3876	T3874	dobj	functions,approach
R843	T3877	T3878	npadvmod	isoform,specific
R844	T3878	T3876	amod	specific,functions
R845	T3879	T3852	punct	.,provides
R846	T3698	T3695	appos	inhibitor,calpastatin
R847	T3881	T3882	amod	Initial,studies
R848	T3882	T3883	nsubj	studies,targeted
R849	T3699	T3698	amod	endogenous,inhibitor
R85	T759	T758	punct	-,calpain
R850	T3884	T3883	dobj	Capn4,targeted
R851	T3885	T3883	prep	based,targeted
R852	T3700	T3698	compound	protein,inhibitor
R853	T3886	T3885	prep	on,based
R854	T3887	T3888	det	the,prediction
R855	T3888	T3886	pobj	prediction,on
R856	T3889	T3890	mark	that,abolish
R857	T3701	T3698	prep	of,inhibitor
R858	T3890	T3888	acl	abolish,prediction
R859	T3702	T3703	nmod	μ,calpain
R86	T760	T745	punct	", ",abolished
R860	T3891	T3890	nsubj	loss,abolish
R861	T3892	T3891	prep	of,loss
R862	T3893	T3894	det	this,subunit
R863	T3894	T3892	pobj	subunit,of
R864	T3703	T3701	pobj	calpain,of
R865	T3895	T3894	compound	calpain,subunit
R866	T3896	T3890	aux	would,abolish
R867	T3704	T3702	punct	-,μ
R868	T3897	T3890	dobj	activity,abolish
R869	T3705	T3702	cc	and,μ
R87	T761	T745	cc	and,abolished
R870	T3898	T3897	prep	of,activity
R871	T3899	T3900	preconj	both,μ
R872	T3706	T3702	conj	m,μ
R873	T3900	T3901	nmod	μ,calpain
R874	T3901	T3898	pobj	calpain,of
R875	T3707	T3703	punct	-,calpain
R876	T3902	T3900	punct	-,μ
R877	T3903	T3900	cc	and,μ
R878	T3904	T3900	conj	m,μ
R879	T3709	T3710	det	an,approach
R88	T762	T745	conj	resulted,abolished
R880	T3905	T3901	punct	-,calpain
R881	T3906	T3883	punct	.,targeted
R882	T3908	T3909	nmod	Capn4,embryos
R883	T3710	T3692	dobj	approach,provides
R884	T3909	T3914	nsubj	embryos,died
R885	T3910	T3908	punct	-,Capn4
R886	T3911	T3908	punct	/,Capn4
R887	T3711	T3710	amod	important,approach
R888	T3912	T3908	punct	-,Capn4
R889	T3913	T3909	amod	murine,embryos
R89	T763	T762	prep	in,resulted
R890	T3712	T3710	prep	for,approach
R891	T3915	T3914	prep	between,died
R892	T3916	T3917	nmod	days,10
R893	T3713	T3714	det	these,efforts
R894	T3917	T3915	pobj	10,between
R895	T3918	T3917	cc	and,10
R896	T3919	T3917	conj	11,10
R897	T3920	T3917	prep	of,10
R898	T3921	T3920	pobj	gestation,of
R899	T3922	T3914	punct	", ",died
R9	T673	T671	pobj	development,for
R90	T764	T765	amod	embryonic,lethality
R900	T3714	T3712	pobj	efforts,for
R901	T3923	T3914	cc	and,died
R902	T3924	T3925	expl	there,was
R903	T3925	T3914	conj	was,died
R904	T3715	T3708	punct	", ",distinguish
R905	T3926	T3927	det	no,activity
R906	T3927	T3925	attr	activity,was
R907	T3928	T3927	amod	detectable,activity
R908	T3716	T3708	nsubj	it,distinguish
R909	T3929	T3930	nmod	μ,calpain
R91	T765	T763	pobj	lethality,in
R910	T3930	T3927	compound	calpain,activity
R911	T3931	T3929	punct	-,μ
R912	T3717	T3708	aux	will,distinguish
R913	T3932	T3929	cc	or,μ
R914	T3933	T3929	conj	m,μ
R915	T3718	T3708	neg	not,distinguish
R916	T3934	T3930	punct	-,calpain
R917	T3935	T3925	prep	in,was
R918	T3936	T3935	pobj	these,in
R919	T3937	T3936	cc	or,these
R92	T766	T762	punct	", ",resulted
R920	T3719	T3720	npadvmod	isoform,specific
R921	T3938	T3939	amod	younger,embryos
R922	T3939	T3936	conj	embryos,these
R923	T3720	T3721	amod	specific,functions
R924	T3940	T3941	punct	[,31
R925	T3941	T3925	parataxis	31,was
R926	T3942	T3941	punct	],31
R927	T3721	T3708	dobj	functions,distinguish
R928	T3943	T3925	punct	.,was
R929	T3945	T3946	nsubjpass	Capn4,cultured
R93	T767	T768	advmod	thereby,suggesting
R930	T3722	T3723	punct	[,27
R931	T3947	T3945	punct	-,Capn4
R932	T3948	T3945	punct	/,Capn4
R933	T3723	T3708	parataxis	27,distinguish
R934	T3949	T3945	punct	-,Capn4
R935	T3950	T3951	amod	murine,fibroblasts
R936	T3724	T3723	nummod	24,27
R937	T3725	T3723	punct	",",27
R938	T3951	T3945	appos	fibroblasts,Capn4
R939	T3726	T3723	nummod	26,27
R94	T768	T762	advcl	suggesting,resulted
R940	T3952	T3951	amod	embryonic,fibroblasts
R941	T3727	T3723	punct	",",27
R942	T3953	T3951	punct	(,fibroblasts
R943	T3954	T3951	appos	MEFs,fibroblasts
R944	T3728	T3723	punct	],27
R945	T3729	T3708	punct	.,distinguish
R946	T3955	T3946	punct	),cultured
R947	T3731	T3732	det	Some,work
R948	T3956	T3946	aux	could,cultured
R949	T3957	T3946	auxpass	be,cultured
R95	T769	T770	amod	essential,roles
R950	T3958	T3946	prep	from,cultured
R951	T3959	T3960	det	these,embryos
R952	T3960	T3958	pobj	embryos,from
R953	T3961	T3946	punct	", ",cultured
R954	T3962	T3963	mark	although,lacked
R955	T3732	T3733	nsubj	work,suggested
R956	T3963	T3946	advcl	lacked,cultured
R957	T3964	T3963	nsubj	they,lacked
R958	T3965	T3963	advmod	also,lacked
R959	T3734	T3733	aux	has,suggested
R96	T770	T768	dobj	roles,suggesting
R960	T3966	T3967	compound	calpain,activity
R961	T3967	T3963	dobj	activity,lacked
R962	T3968	T3969	mark	as,assessed
R963	T3969	T3963	advcl	assessed,lacked
R964	T3735	T3736	npadvmod	isoform,specific
R965	T3970	T3969	prep	by,assessed
R966	T3971	T3972	compound	casein,zymography
R967	T3972	T3970	pobj	zymography,by
R968	T3736	T3737	amod	specific,roles
R969	T3973	T3970	cc	or,by
R97	T771	T770	prep	for,roles
R970	T3974	T3970	conj	by,by
R971	T3737	T3733	dobj	roles,suggested
R972	T3975	T3976	det	the,formation
R973	T3976	T3974	pobj	formation,by
R974	T3738	T3737	punct	", ",roles
R975	T3977	T3976	prep	of,formation
R976	T3978	T3979	amod	characteristic,products
R977	T3979	T3977	pobj	products,of
R978	T3739	T3740	amod	such,as
R979	T3980	T3981	compound	spectrin,breakdown
R98	T772	T771	pobj	one,for
R980	T3981	T3979	compound	breakdown,products
R981	T3740	T3737	prep	as,roles
R982	T3741	T3742	det	a,role
R983	T3982	T3963	punct	", ",lacked
R984	T3983	T3963	cc	and,lacked
R985	T3742	T3740	pobj	role,as
R986	T3984	T3985	nsubj	they,displayed
R987	T3985	T3963	conj	displayed,lacked
R988	T3986	T3987	compound	migration,defects
R989	T3743	T3742	prep	for,role
R99	T773	T772	cc	or,one
R990	T3987	T3985	dobj	defects,displayed
R991	T3988	T3987	amod	consistent,defects
R992	T3989	T3988	prep	with,consistent
R993	T3744	T3745	compound	m,calpain
R994	T3990	T3991	det	a,role
R995	T3991	T3989	pobj	role,with
R996	T3745	T3743	pobj	calpain,for
R997	T3992	T3991	prep	for,role
R998	T3993	T3992	pobj	calpain,for
R999	T3994	T3991	prep	in,role

PMC:1382200 JSONTXT 7 Projects

Annnotations TAB TSV DIC JSON TextAE

PMC:1382200 JSON TXT 7 Projects