PMC:1435744 / 40427-48657 JSON TXT 4 Projects

Annnotations TAB TSV DIC JSON TextAE

Id	Subject	Object	Predicate	Lexical cue
T23217	9-18	NN	denotes	Isolation
T23218	19-21	IN	denotes	of
T23219	22-27	NN	denotes	mouse
T23221	28-30	NN	denotes	mr
T23223	30-31	HYPH	denotes	-
T23222	31-32	NN	denotes	s
T23220	33-37	NN	denotes	cDNA
T23224	37-184	sentence	denotes	We used the bioinformatics method Digital Differential Display (NCBI, UniGene) to screen novel mouse genes expressed preferentially in the retina.
T23225	38-40	PRP	denotes	We
T23226	41-45	VBD	denotes	used
T23227	46-49	DT	denotes	the
T23229	50-64	NN	denotes	bioinformatics
T23228	65-71	NN	denotes	method
T23230	72-79	NNP	denotes	Digital
T23232	80-92	NNP	denotes	Differential
T23231	93-100	NNP	denotes	Display
T23233	101-102	-LRB-	denotes	(
T23234	102-106	NNP	denotes	NCBI
T23235	106-108	,	denotes	,
T23236	108-115	NNP	denotes	UniGene
T23237	115-116	-RRB-	denotes	)
T23238	117-119	TO	denotes	to
T23239	120-126	VB	denotes	screen
T23240	127-132	JJ	denotes	novel
T23242	133-138	NN	denotes	mouse
T23241	139-144	NNS	denotes	genes
T23243	145-154	VBN	denotes	expressed
T23244	155-169	RB	denotes	preferentially
T23245	170-172	IN	denotes	in
T23246	173-176	DT	denotes	the
T23247	177-183	NN	denotes	retina
T23248	183-184	.	denotes	.
T23249	184-267	sentence	denotes	Some of the clusters in the UniGene database were mainly from mouse retinal cDNAs.
T23250	185-189	DT	denotes	Some
T23252	190-192	IN	denotes	of
T23253	193-196	DT	denotes	the
T23254	197-205	NNS	denotes	clusters
T23255	206-208	IN	denotes	in
T23256	209-212	DT	denotes	the
T23258	213-220	NNP	denotes	UniGene
T23257	221-229	NN	denotes	database
T23251	230-234	VBD	denotes	were
T23259	235-241	RB	denotes	mainly
T23260	242-246	IN	denotes	from
T23261	247-252	NN	denotes	mouse
T23263	253-260	JJ	denotes	retinal
T23262	261-266	NNS	denotes	cDNAs
T23264	266-267	.	denotes	.
T23265	267-365	sentence	denotes	One clone in these clusters (#Mm. 246385) has a weak homology with the polyhomeotic family genes.
T23266	268-271	CD	denotes	One
T23267	272-277	NN	denotes	clone
T23269	278-280	IN	denotes	in
T23270	281-286	DT	denotes	these
T23271	287-295	NNS	denotes	clusters
T23272	296-297	-LRB-	denotes	(
T23274	297-298	SYM	denotes	#
T23273	298-301	NNP	denotes	Mm.
T23275	302-308	CD	denotes	246385
T23276	308-309	-RRB-	denotes	)
T23268	310-313	VBZ	denotes	has
T23277	314-315	DT	denotes	a
T23279	316-320	JJ	denotes	weak
T23278	321-329	NN	denotes	homology
T23280	330-334	IN	denotes	with
T23281	335-338	DT	denotes	the
T23283	339-351	JJ	denotes	polyhomeotic
T23284	352-358	NN	denotes	family
T23282	359-364	NNS	denotes	genes
T23285	364-365	.	denotes	.
T23286	365-484	sentence	denotes	A 735-bp cDNA fragment of this clone, encoding amino acids 140–384 was amplified by RT-PCR from mouse P0 retinal cDNA.
T23287	366-367	DT	denotes	A
T23289	368-371	CD	denotes	735
T23291	371-372	HYPH	denotes	-
T23290	372-374	NN	denotes	bp
T23292	375-379	NN	denotes	cDNA
T23288	380-388	NN	denotes	fragment
T23294	389-391	IN	denotes	of
T23295	392-396	DT	denotes	this
T23296	397-402	NN	denotes	clone
T23297	402-404	,	denotes	,
T23298	404-412	VBG	denotes	encoding
T23299	413-418	NN	denotes	amino
T23301	419-424	NNS	denotes	acids
T23300	425-428	CD	denotes	140
T23302	428-429	SYM	denotes	–
T23303	429-432	CD	denotes	384
T23304	433-436	VBD	denotes	was
T23293	437-446	VBN	denotes	amplified
T23305	447-449	IN	denotes	by
T23306	450-452	NN	denotes	RT
T23308	452-453	HYPH	denotes	-
T23307	453-456	NN	denotes	PCR
T23309	457-461	IN	denotes	from
T23310	462-467	NN	denotes	mouse
T23312	468-470	NN	denotes	P0
T23313	471-478	JJ	denotes	retinal
T23311	479-483	NN	denotes	cDNA
T23314	483-484	.	denotes	.
T23315	484-593	sentence	denotes	This fragment was used as the probe for library screening, in situ hybridization and Northern hybridization.
T23316	485-489	DT	denotes	This
T23317	490-498	NN	denotes	fragment
T23319	499-502	VBD	denotes	was
T23318	503-507	VBN	denotes	used
T23320	508-510	IN	denotes	as
T23321	511-514	DT	denotes	the
T23322	515-520	NN	denotes	probe
T23323	521-524	IN	denotes	for
T23324	525-532	NN	denotes	library
T23325	533-542	NN	denotes	screening
T23326	542-544	,	denotes	,
T23327	544-546	FW	denotes	in
T23328	547-551	FW	denotes	situ
T23329	552-565	NN	denotes	hybridization
T23330	566-569	CC	denotes	and
T23331	570-578	NNP	denotes	Northern
T23332	579-592	NN	denotes	hybridization
T23333	592-593	.	denotes	.
T23334	593-673	sentence	denotes	A mouse P0-P3 retinal cDNA library was screened using this mouse cDNA fragment.
T23335	594-595	DT	denotes	A
T23337	596-601	NN	denotes	mouse
T23338	602-604	NN	denotes	P0
T23339	604-605	SYM	denotes	-
T23340	605-607	NN	denotes	P3
T23341	608-615	JJ	denotes	retinal
T23342	616-620	NN	denotes	cDNA
T23336	621-628	NN	denotes	library
T23344	629-632	VBD	denotes	was
T23343	633-641	VBN	denotes	screened
T23345	642-647	VBG	denotes	using
T23346	648-652	DT	denotes	this
T23348	653-658	NN	denotes	mouse
T23349	659-663	NN	denotes	cDNA
T23347	664-672	NN	denotes	fragment
T23350	672-673	.	denotes	.
T23351	673-797	sentence	denotes	Positive bacteriophage clones were isolated and the full-length mr-s fragment was inserted into pBluescriptII (Stratagene).
T23352	674-682	JJ	denotes	Positive
T23354	683-696	NN	denotes	bacteriophage
T23353	697-703	NNS	denotes	clones
T23356	704-708	VBD	denotes	were
T23355	709-717	VBN	denotes	isolated
T23357	718-721	CC	denotes	and
T23358	722-725	DT	denotes	the
T23360	726-730	JJ	denotes	full
T23362	730-731	HYPH	denotes	-
T23361	731-737	NN	denotes	length
T23363	738-740	NN	denotes	mr
T23365	740-741	HYPH	denotes	-
T23364	741-742	NN	denotes	s
T23359	743-751	NN	denotes	fragment
T23367	752-755	VBD	denotes	was
T23366	756-764	VBN	denotes	inserted
T23368	765-769	IN	denotes	into
T23369	770-783	NNP	denotes	pBluescriptII
T23370	784-785	-LRB-	denotes	(
T23371	785-795	NNP	denotes	Stratagene
T23372	795-796	-RRB-	denotes	)
T23373	796-797	.	denotes	.
T23374	797-874	sentence	denotes	DNA sequencing was performed on both strands by the cycle sequencing method.
T23375	798-801	NN	denotes	DNA
T23376	802-812	NN	denotes	sequencing
T23378	813-816	VBD	denotes	was
T23377	817-826	VBN	denotes	performed
T23379	827-829	IN	denotes	on
T23380	830-834	DT	denotes	both
T23381	835-842	NNS	denotes	strands
T23382	843-845	IN	denotes	by
T23383	846-849	DT	denotes	the
T23385	850-855	NN	denotes	cycle
T23386	856-866	NN	denotes	sequencing
T23384	867-873	NN	denotes	method
T23387	873-874	.	denotes	.
T23388	874-997	sentence	denotes	The nucleotide sequence for mr-s gene has been deposited in the GenBank database under GenBank Accession Number #AY458844.
T23389	875-878	DT	denotes	The
T23391	879-889	NN	denotes	nucleotide
T23390	890-898	NN	denotes	sequence
T23393	899-902	IN	denotes	for
T23394	903-905	NN	denotes	mr
T23396	905-906	HYPH	denotes	-
T23395	906-907	NN	denotes	s
T23397	908-912	NN	denotes	gene
T23398	913-916	VBZ	denotes	has
T23399	917-921	VBN	denotes	been
T23392	922-931	VBN	denotes	deposited
T23400	932-934	IN	denotes	in
T23401	935-938	DT	denotes	the
T23403	939-946	NNP	denotes	GenBank
T23402	947-955	NN	denotes	database
T23404	956-961	IN	denotes	under
T23405	962-969	NNP	denotes	GenBank
T23407	970-979	NNP	denotes	Accession
T23408	980-986	NNP	denotes	Number
T23409	987-988	SYM	denotes	#
T23406	988-996	NN	denotes	AY458844
T23410	996-997	.	denotes	.
T23508	999-1001	FW	denotes	In
T23509	1002-1006	FW	denotes	situ
T23510	1007-1020	NN	denotes	hybridization
T23511	1020-1120	sentence	denotes	The 735-bp cDNA fragment of mr-s amplified by RT-PCR was used as a probe for in situ hybridization.
T23512	1021-1024	DT	denotes	The
T23514	1025-1028	CD	denotes	735
T23516	1028-1029	HYPH	denotes	-
T23515	1029-1031	NN	denotes	bp
T23517	1032-1036	NN	denotes	cDNA
T23513	1037-1045	NN	denotes	fragment
T23519	1046-1048	IN	denotes	of
T23520	1049-1051	NN	denotes	mr
T23522	1051-1052	HYPH	denotes	-
T23521	1052-1053	NN	denotes	s
T23523	1054-1063	VBN	denotes	amplified
T23524	1064-1066	IN	denotes	by
T23525	1067-1069	NN	denotes	RT
T23527	1069-1070	HYPH	denotes	-
T23526	1070-1073	NN	denotes	PCR
T23528	1074-1077	VBD	denotes	was
T23518	1078-1082	VBN	denotes	used
T23529	1083-1085	IN	denotes	as
T23530	1086-1087	DT	denotes	a
T23531	1088-1093	NN	denotes	probe
T23532	1094-1097	IN	denotes	for
T23533	1098-1100	FW	denotes	in
T23534	1101-1105	FW	denotes	situ
T23535	1106-1119	NN	denotes	hybridization
T23536	1119-1120	.	denotes	.
T23537	1120-1186	sentence	denotes	In situ hybridization was performed as described previously [49].
T23538	1121-1123	FW	denotes	In
T23539	1124-1128	FW	denotes	situ
T23540	1129-1142	NN	denotes	hybridization
T23542	1143-1146	VBD	denotes	was
T23541	1147-1156	VBN	denotes	performed
T23543	1157-1159	IN	denotes	as
T23544	1160-1169	VBN	denotes	described
T23545	1170-1180	RB	denotes	previously
T23546	1181-1182	-LRB-	denotes	[
T23547	1182-1184	CD	denotes	49
T23548	1184-1185	-RRB-	denotes	]
T23549	1185-1186	.	denotes	.
T23802	1188-1192	NN	denotes	Cell
T23803	1193-1200	NN	denotes	culture
T23804	1201-1204	CC	denotes	and
T23805	1205-1217	NN	denotes	transfection
T23806	1217-1399	sentence	denotes	HEK293T cells were maintained at 37°C in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (Sigma), 100 IU/ml penicillin and 100 μg/ml streptomycin.
T23807	1218-1225	NN	denotes	HEK293T
T23808	1226-1231	NNS	denotes	cells
T23810	1232-1236	VBD	denotes	were
T23809	1237-1247	VBN	denotes	maintained
T23811	1248-1250	IN	denotes	at
T23812	1251-1253	CD	denotes	37
T23813	1253-1255	NN	denotes	°C
T23814	1256-1258	IN	denotes	in
T23815	1259-1267	NNP	denotes	Dulbecco
T23817	1267-1269	POS	denotes	's
T23818	1270-1278	VBN	denotes	modified
T23819	1279-1284	NNP	denotes	Eagle
T23820	1284-1286	POS	denotes	's
T23816	1287-1293	NN	denotes	medium
T23821	1294-1295	-LRB-	denotes	(
T23822	1295-1299	NN	denotes	DMEM
T23823	1299-1300	-RRB-	denotes	)
T23824	1301-1313	VBN	denotes	supplemented
T23825	1314-1318	IN	denotes	with
T23826	1319-1321	CD	denotes	10
T23827	1321-1322	NN	denotes	%
T23829	1323-1328	JJ	denotes	fetal
T23830	1329-1335	JJ	denotes	bovine
T23828	1336-1341	NN	denotes	serum
T23831	1342-1343	-LRB-	denotes	(
T23832	1343-1348	NNP	denotes	Sigma
T23833	1348-1349	-RRB-	denotes	)
T23834	1349-1351	,	denotes	,
T23835	1351-1354	CD	denotes	100
T23836	1355-1357	NN	denotes	IU
T23838	1357-1358	SYM	denotes	/
T23839	1358-1360	NN	denotes	ml
T23837	1361-1371	NN	denotes	penicillin
T23840	1372-1375	CC	denotes	and
T23841	1376-1379	CD	denotes	100
T23842	1380-1382	NN	denotes	μg
T23844	1382-1383	SYM	denotes	/
T23845	1383-1385	NN	denotes	ml
T23843	1386-1398	NN	denotes	streptomycin
T23846	1398-1399	.	denotes	.
T23847	1399-1527	sentence	denotes	Transient transfection of HEK293T cells was carried out using calcium phosphate method or Fugene6 transfection reagent (Roche).
T23848	1400-1409	JJ	denotes	Transient
T23849	1410-1422	NN	denotes	transfection
T23851	1423-1425	IN	denotes	of
T23852	1426-1433	NN	denotes	HEK293T
T23853	1434-1439	NNS	denotes	cells
T23854	1440-1443	VBD	denotes	was
T23850	1444-1451	VBN	denotes	carried
T23855	1452-1455	RP	denotes	out
T23856	1456-1461	VBG	denotes	using
T23857	1462-1469	NN	denotes	calcium
T23858	1470-1479	NN	denotes	phosphate
T23859	1480-1486	NN	denotes	method
T23860	1487-1489	CC	denotes	or
T23861	1490-1497	NN	denotes	Fugene6
T23863	1498-1510	NN	denotes	transfection
T23862	1511-1518	NN	denotes	reagent
T23864	1519-1520	-LRB-	denotes	(
T23865	1520-1525	NNP	denotes	Roche
T23866	1525-1526	-RRB-	denotes	)
T23867	1526-1527	.	denotes	.
T23868	1527-1686	sentence	denotes	Y79 retinoblastoma cells were maintained in Iscove's modified Dulbecco's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 20% FBS.
T23869	1528-1531	NN	denotes	Y79
T23871	1532-1546	NN	denotes	retinoblastoma
T23870	1547-1552	NNS	denotes	cells
T23873	1553-1557	VBD	denotes	were
T23872	1558-1568	VBN	denotes	maintained
T23874	1569-1571	IN	denotes	in
T23875	1572-1578	NNP	denotes	Iscove
T23877	1578-1580	POS	denotes	's
T23878	1581-1589	VBN	denotes	modified
T23879	1590-1598	NNP	denotes	Dulbecco
T23880	1598-1600	POS	denotes	's
T23876	1601-1607	NN	denotes	medium
T23881	1608-1612	IN	denotes	with
T23882	1613-1614	CD	denotes	4
T23883	1615-1617	NN	denotes	mM
T23885	1618-1619	NN	denotes	L
T23886	1619-1620	HYPH	denotes	-
T23884	1620-1629	NN	denotes	glutamine
T23887	1630-1638	VBN	denotes	adjusted
T23888	1639-1641	TO	denotes	to
T23889	1642-1649	VB	denotes	contain
T23890	1650-1653	CD	denotes	1.5
T23891	1654-1655	NN	denotes	g
T23893	1655-1656	SYM	denotes	/
T23894	1656-1657	NN	denotes	L
T23895	1658-1664	NN	denotes	sodium
T23892	1665-1676	NN	denotes	bicarbonate
T23896	1676-1678	,	denotes	,
T23897	1678-1680	CD	denotes	20
T23898	1680-1681	NN	denotes	%
T23899	1682-1685	NN	denotes	FBS
T23900	1685-1686	.	denotes	.
T23901	1686-1752	sentence	denotes	Transient transfection was carried out using TransIt LT1 (Mirus).
T23902	1687-1696	JJ	denotes	Transient
T23903	1697-1709	NN	denotes	transfection
T23905	1710-1713	VBD	denotes	was
T23904	1714-1721	VBN	denotes	carried
T23906	1722-1725	RP	denotes	out
T23907	1726-1731	VBG	denotes	using
T23908	1732-1739	NN	denotes	TransIt
T23909	1740-1743	NN	denotes	LT1
T23910	1744-1745	-LRB-	denotes	(
T23911	1745-1750	NNP	denotes	Mirus
T23912	1750-1751	-RRB-	denotes	)
T23913	1751-1752	.	denotes	.
T24169	1754-1762	NNP	denotes	Northern
T24171	1762-1763	HYPH	denotes	-
T24170	1763-1767	NN	denotes	blot
T24172	1768-1776	NN	denotes	analysis
T24173	1776-1852	sentence	denotes	RNA was extracted from the tissues of adult mice using Trizol (Invitrogen).
T24174	1777-1780	NN	denotes	RNA
T24176	1781-1784	VBD	denotes	was
T24175	1785-1794	VBN	denotes	extracted
T24177	1795-1799	IN	denotes	from
T24178	1800-1803	DT	denotes	the
T24179	1804-1811	NNS	denotes	tissues
T24180	1812-1814	IN	denotes	of
T24181	1815-1820	JJ	denotes	adult
T24182	1821-1825	NNS	denotes	mice
T24183	1826-1831	VBG	denotes	using
T24184	1832-1838	NNP	denotes	Trizol
T24185	1839-1840	-LRB-	denotes	(
T24186	1840-1850	NNP	denotes	Invitrogen
T24187	1850-1851	-RRB-	denotes	)
T24188	1851-1852	.	denotes	.
T24189	1852-1989	sentence	denotes	A 5 μg of total RNA was electrophoresed in a 1.0% agarose-formaldehyde gel and transferred to a nylon membrane (Zeta-Probe GT, Bio-Rad).
T24190	1853-1854	DT	denotes	A
T24192	1855-1856	CD	denotes	5
T24191	1857-1859	NN	denotes	μg
T24194	1860-1862	IN	denotes	of
T24195	1863-1868	JJ	denotes	total
T24196	1869-1872	NN	denotes	RNA
T24197	1873-1876	VBD	denotes	was
T24193	1877-1892	VBN	denotes	electrophoresed
T24198	1893-1895	IN	denotes	in
T24199	1896-1897	DT	denotes	a
T24201	1898-1901	CD	denotes	1.0
T24202	1901-1902	NN	denotes	%
T24203	1903-1910	NN	denotes	agarose
T24205	1910-1911	HYPH	denotes	-
T24204	1911-1923	NN	denotes	formaldehyde
T24200	1924-1927	NN	denotes	gel
T24206	1928-1931	CC	denotes	and
T24207	1932-1943	VBN	denotes	transferred
T24208	1944-1946	IN	denotes	to
T24209	1947-1948	DT	denotes	a
T24211	1949-1954	NN	denotes	nylon
T24210	1955-1963	NN	denotes	membrane
T24212	1964-1965	-LRB-	denotes	(
T24214	1965-1969	NNP	denotes	Zeta
T24216	1969-1970	HYPH	denotes	-
T24215	1970-1975	NNP	denotes	Probe
T24213	1976-1978	NNP	denotes	GT
T24217	1978-1980	,	denotes	,
T24218	1980-1983	NNP	denotes	Bio
T24220	1983-1984	HYPH	denotes	-
T24219	1984-1987	NNP	denotes	Rad
T24221	1987-1988	-RRB-	denotes	)
T24222	1988-1989	.	denotes	.
T24223	1989-2085	sentence	denotes	The 735-bp cDNA fragment encoding the SAM domain of mr-s was used as a probe for hybridization.
T24224	1990-1993	DT	denotes	The
T24226	1994-1997	CD	denotes	735
T24228	1997-1998	HYPH	denotes	-
T24227	1998-2000	NN	denotes	bp
T24229	2001-2005	NN	denotes	cDNA
T24225	2006-2014	NN	denotes	fragment
T24231	2015-2023	VBG	denotes	encoding
T24232	2024-2027	DT	denotes	the
T24234	2028-2031	NN	denotes	SAM
T24233	2032-2038	NN	denotes	domain
T24235	2039-2041	IN	denotes	of
T24236	2042-2044	NN	denotes	mr
T24238	2044-2045	HYPH	denotes	-
T24237	2045-2046	NN	denotes	s
T24239	2047-2050	VBD	denotes	was
T24230	2051-2055	VBN	denotes	used
T24240	2056-2058	IN	denotes	as
T24241	2059-2060	DT	denotes	a
T24242	2061-2066	NN	denotes	probe
T24243	2067-2070	IN	denotes	for
T24244	2071-2084	NN	denotes	hybridization
T24245	2084-2085	.	denotes	.
T24246	2085-2155	sentence	denotes	Hybridization was performed according to the manufacturer's protocol.
T24247	2086-2099	NN	denotes	Hybridization
T24249	2100-2103	VBD	denotes	was
T24248	2104-2113	VBN	denotes	performed
T24250	2114-2123	VBG	denotes	according
T24251	2124-2126	IN	denotes	to
T24252	2127-2130	DT	denotes	the
T24253	2131-2143	NN	denotes	manufacturer
T24255	2143-2145	POS	denotes	's
T24254	2146-2154	NN	denotes	protocol
T24256	2154-2155	.	denotes	.
T24257	2155-2295	sentence	denotes	Washes with increasing stringency were performed, the last being at 50°C in 0.1× standard saline citrate/0.1% sodium dodecyl sulfate (SDS).
T24258	2156-2162	NNS	denotes	Washes
T24260	2163-2167	IN	denotes	with
T24261	2168-2178	VBG	denotes	increasing
T24262	2179-2189	NN	denotes	stringency
T24263	2190-2194	VBD	denotes	were
T24259	2195-2204	VBN	denotes	performed
T24264	2204-2206	,	denotes	,
T24265	2206-2209	DT	denotes	the
T24266	2210-2214	JJ	denotes	last
T24267	2215-2220	VBG	denotes	being
T24268	2221-2223	IN	denotes	at
T24269	2224-2226	CD	denotes	50
T24270	2226-2228	NN	denotes	°C
T24271	2229-2231	IN	denotes	in
T24272	2232-2235	CD	denotes	0.1
T24274	2235-2236	SYM	denotes	×
T24275	2237-2245	JJ	denotes	standard
T24276	2246-2252	NN	denotes	saline
T24273	2253-2260	NN	denotes	citrate
T24277	2260-2261	SYM	denotes	/
T24279	2261-2264	CD	denotes	0.1
T24278	2264-2265	NN	denotes	%
T24280	2266-2272	NN	denotes	sodium
T24282	2273-2280	NN	denotes	dodecyl
T24281	2281-2288	NN	denotes	sulfate
T24283	2289-2290	-LRB-	denotes	(
T24284	2290-2293	NN	denotes	SDS
T24285	2293-2294	-RRB-	denotes	)
T24286	2294-2295	.	denotes	.
T24536	2297-2299	NN	denotes	RT
T24538	2299-2300	HYPH	denotes	-
T24537	2300-2303	NN	denotes	PCR
T24539	2304-2312	NN	denotes	analysis
T24540	2312-2366	sentence	denotes	Total RNA was isolated from each tissue using Trizol.
T24541	2313-2318	JJ	denotes	Total
T24542	2319-2322	NN	denotes	RNA
T24544	2323-2326	VBD	denotes	was
T24543	2327-2335	VBN	denotes	isolated
T24545	2336-2340	IN	denotes	from
T24546	2341-2345	DT	denotes	each
T24547	2346-2352	NN	denotes	tissue
T24548	2353-2358	VBG	denotes	using
T24549	2359-2365	NN	denotes	Trizol
T24550	2365-2366	.	denotes	.
T24551	2366-2444	sentence	denotes	A 1 μg of total RNA was reverse transcribed using SuperscriptII (Invitrogen).
T24552	2367-2368	DT	denotes	A
T24554	2369-2370	CD	denotes	1
T24553	2371-2373	NN	denotes	μg
T24556	2374-2376	IN	denotes	of
T24557	2377-2382	JJ	denotes	total
T24558	2383-2386	NN	denotes	RNA
T24559	2387-2390	VBD	denotes	was
T24560	2391-2398	JJ	denotes	reverse
T24555	2399-2410	VBN	denotes	transcribed
T24561	2411-2416	VBG	denotes	using
T24562	2417-2430	NNP	denotes	SuperscriptII
T24563	2431-2432	-LRB-	denotes	(
T24564	2432-2442	NNP	denotes	Invitrogen
T24565	2442-2443	-RRB-	denotes	)
T24566	2443-2444	.	denotes	.
T24567	2444-2585	sentence	denotes	RT-PCR primers, which span introns, for detection of mr-s cDNA were 5'-TGTCCAGCCCAGCCAACCCAAGGAGACGACA-3' and 5'-TGTGGTCTCCTCATCAGTGAAGA-3'.
T24568	2445-2447	NN	denotes	RT
T24570	2447-2448	HYPH	denotes	-
T24569	2448-2451	NN	denotes	PCR
T24571	2452-2459	NNS	denotes	primers
T24573	2459-2461	,	denotes	,
T24574	2461-2466	WDT	denotes	which
T24575	2467-2471	VBP	denotes	span
T24576	2472-2479	NNS	denotes	introns
T24577	2479-2481	,	denotes	,
T24578	2481-2484	IN	denotes	for
T24579	2485-2494	NN	denotes	detection
T24580	2495-2497	IN	denotes	of
T24581	2498-2500	NN	denotes	mr
T24583	2500-2501	HYPH	denotes	-
T24582	2501-2502	NN	denotes	s
T24584	2503-2507	NN	denotes	cDNA
T24572	2508-2512	VBD	denotes	were
T24585	2513-2514	CD	denotes	5
T24587	2514-2515	SYM	denotes	'
T24588	2515-2516	HYPH	denotes	-
T24586	2516-2547	NN	denotes	TGTCCAGCCCAGCCAACCCAAGGAGACGACA
T24589	2547-2548	HYPH	denotes	-
T24590	2548-2549	CD	denotes	3
T24591	2549-2550	SYM	denotes	'
T24592	2551-2554	CC	denotes	and
T24593	2555-2556	CD	denotes	5
T24595	2556-2557	SYM	denotes	'
T24596	2557-2558	HYPH	denotes	-
T24594	2558-2581	NN	denotes	TGTGGTCTCCTCATCAGTGAAGA
T24597	2581-2582	HYPH	denotes	-
T24598	2582-2583	CD	denotes	3
T24599	2583-2584	SYM	denotes	'
T24600	2584-2585	.	denotes	.
T24601	2585-2669	sentence	denotes	Product size was 292 bp (positions 965–1256 of Genbank Accession Number #AY458844).
T24602	2586-2593	NN	denotes	Product
T24603	2594-2598	NN	denotes	size
T24604	2599-2602	VBD	denotes	was
T24605	2603-2606	CD	denotes	292
T24606	2607-2609	NN	denotes	bp
T24607	2610-2611	-LRB-	denotes	(
T24609	2611-2620	NNS	denotes	positions
T24608	2621-2624	CD	denotes	965
T24610	2624-2625	SYM	denotes	–
T24611	2625-2629	CD	denotes	1256
T24612	2630-2632	IN	denotes	of
T24613	2633-2640	NNP	denotes	Genbank
T24615	2641-2650	NNP	denotes	Accession
T24616	2651-2657	NNP	denotes	Number
T24617	2658-2659	SYM	denotes	#
T24614	2659-2667	NN	denotes	AY458844
T24618	2667-2668	-RRB-	denotes	)
T24619	2668-2669	.	denotes	.
T24620	2669-2854	sentence	denotes	Primer pairs for mouse G3PDH were 5'-ACCACAGTCCATGCCATCAC-3' and 5'-TCCACCACCCTGTTGCTGTA- 3' which amplified a 452-bp product (positions 587–1038 of Genbank Accession Number #BC85275).
T24621	2670-2676	NN	denotes	Primer
T24622	2677-2682	NNS	denotes	pairs
T24624	2683-2686	IN	denotes	for
T24625	2687-2692	NN	denotes	mouse
T24626	2693-2698	NN	denotes	G3PDH
T24623	2699-2703	VBD	denotes	were
T24627	2704-2705	CD	denotes	5
T24629	2705-2706	SYM	denotes	'
T24630	2706-2707	HYPH	denotes	-
T24628	2707-2727	NN	denotes	ACCACAGTCCATGCCATCAC
T24631	2727-2728	HYPH	denotes	-
T24632	2728-2729	CD	denotes	3
T24633	2729-2730	SYM	denotes	'
T24634	2731-2734	CC	denotes	and
T24635	2735-2736	CD	denotes	5
T24637	2736-2737	SYM	denotes	'
T24638	2737-2738	HYPH	denotes	-
T24636	2738-2758	NN	denotes	TCCACCACCCTGTTGCTGTA
T24639	2758-2759	HYPH	denotes	-
T24640	2760-2761	CD	denotes	3
T24641	2761-2762	SYM	denotes	'
T24642	2763-2768	WDT	denotes	which
T24643	2769-2778	VBD	denotes	amplified
T24644	2779-2780	DT	denotes	a
T24646	2781-2784	CD	denotes	452
T24648	2784-2785	HYPH	denotes	-
T24647	2785-2787	NN	denotes	bp
T24645	2788-2795	NN	denotes	product
T24649	2796-2797	-LRB-	denotes	(
T24651	2797-2806	NNS	denotes	positions
T24650	2807-2810	CD	denotes	587
T24652	2810-2811	SYM	denotes	–
T24653	2811-2815	CD	denotes	1038
T24654	2816-2818	IN	denotes	of
T24655	2819-2826	NNP	denotes	Genbank
T24657	2827-2836	NN	denotes	Accession
T24658	2837-2843	NN	denotes	Number
T24659	2844-2845	SYM	denotes	#
T24656	2845-2852	NN	denotes	BC85275
T24660	2852-2853	-RRB-	denotes	)
T24661	2853-2854	.	denotes	.
T25116	2856-2861	NN	denotes	Yeast
T25118	2862-2865	CD	denotes	two
T25120	2865-2866	HYPH	denotes	-
T25119	2866-2872	NN	denotes	hybrid
T25117	2873-2882	NN	denotes	screening
T25121	2883-2886	CC	denotes	and
T25122	2887-2891	NN	denotes	GAL4
T25123	2892-2897	NN	denotes	assay
T25124	2897-3039	sentence	denotes	We carried out yeast two-hybrid experiments using the MATCHMAKER GAL4 two-hybrid system 3 (BD Bioscience) as recommended by the manufacturer.
T25125	2898-2900	PRP	denotes	We
T25126	2901-2908	VBD	denotes	carried
T25127	2909-2912	RP	denotes	out
T25128	2913-2918	NN	denotes	yeast
T25130	2919-2922	CD	denotes	two
T25132	2922-2923	HYPH	denotes	-
T25131	2923-2929	NN	denotes	hybrid
T25129	2930-2941	NNS	denotes	experiments
T25133	2942-2947	VBG	denotes	using
T25134	2948-2951	DT	denotes	the
T25136	2952-2962	NNP	denotes	MATCHMAKER
T25137	2963-2967	NN	denotes	GAL4
T25138	2968-2971	CD	denotes	two
T25140	2971-2972	HYPH	denotes	-
T25139	2972-2978	NN	denotes	hybrid
T25135	2979-2985	NN	denotes	system
T25141	2986-2987	CD	denotes	3
T25142	2988-2989	-LRB-	denotes	(
T25144	2989-2991	NNP	denotes	BD
T25143	2992-3002	NNP	denotes	Bioscience
T25145	3002-3003	-RRB-	denotes	)
T25146	3004-3006	IN	denotes	as
T25147	3007-3018	VBN	denotes	recommended
T25148	3019-3021	IN	denotes	by
T25149	3022-3025	DT	denotes	the
T25150	3026-3038	NN	denotes	manufacturer
T25151	3038-3039	.	denotes	.
T25152	3039-3176	sentence	denotes	We cloned the full-length mr-s into the pGBKT7 vector and used it to screen a library of mouse P0-P3 retinal cDNAs in the pGADT7 vector.
T25153	3040-3042	PRP	denotes	We
T25154	3043-3049	VBD	denotes	cloned
T25155	3050-3053	DT	denotes	the
T25157	3054-3058	JJ	denotes	full
T25159	3058-3059	HYPH	denotes	-
T25158	3059-3065	NN	denotes	length
T25160	3066-3068	NN	denotes	mr
T25161	3068-3069	HYPH	denotes	-
T25156	3069-3070	NN	denotes	s
T25162	3071-3075	IN	denotes	into
T25163	3076-3079	DT	denotes	the
T25165	3080-3086	NN	denotes	pGBKT7
T25164	3087-3093	NN	denotes	vector
T25166	3094-3097	CC	denotes	and
T25167	3098-3102	VBD	denotes	used
T25168	3103-3105	PRP	denotes	it
T25169	3106-3108	TO	denotes	to
T25170	3109-3115	VB	denotes	screen
T25171	3116-3117	DT	denotes	a
T25172	3118-3125	NN	denotes	library
T25173	3126-3128	IN	denotes	of
T25174	3129-3134	NN	denotes	mouse
T25176	3135-3137	NN	denotes	P0
T25177	3137-3138	SYM	denotes	-
T25178	3138-3140	NN	denotes	P3
T25179	3141-3148	JJ	denotes	retinal
T25175	3149-3154	NNS	denotes	cDNAs
T25180	3155-3157	IN	denotes	in
T25181	3158-3161	DT	denotes	the
T25183	3162-3168	NN	denotes	pGADT7
T25182	3169-3175	NN	denotes	vector
T25184	3175-3176	.	denotes	.
T25185	3176-3301	sentence	denotes	Transformants that conferred growth were picked, isolated and re-introduced with the bait into AH109 to confirm interaction.
T25186	3177-3190	NNS	denotes	Transformants
T25188	3191-3195	WDT	denotes	that
T25189	3196-3205	VBD	denotes	conferred
T25190	3206-3212	NN	denotes	growth
T25191	3213-3217	VBD	denotes	were
T25187	3218-3224	VBN	denotes	picked
T25192	3224-3226	,	denotes	,
T25193	3226-3234	VBN	denotes	isolated
T25194	3235-3238	CC	denotes	and
T25195	3239-3252	VBN	denotes	re-introduced
T25196	3253-3257	IN	denotes	with
T25197	3258-3261	DT	denotes	the
T25198	3262-3266	NN	denotes	bait
T25199	3267-3271	IN	denotes	into
T25200	3272-3277	NN	denotes	AH109
T25201	3278-3280	TO	denotes	to
T25202	3281-3288	VB	denotes	confirm
T25203	3289-3300	NN	denotes	interaction
T25204	3300-3301	.	denotes	.
T25205	3301-3387	sentence	denotes	Plasmid DNA was isolated from yeast using RPM Yeast Plasmid Isolation Kit (Qbiogene).
T25206	3302-3309	NN	denotes	Plasmid
T25207	3310-3313	NN	denotes	DNA
T25209	3314-3317	VBD	denotes	was
T25208	3318-3326	VBN	denotes	isolated
T25210	3327-3331	IN	denotes	from
T25211	3332-3337	NN	denotes	yeast
T25212	3338-3343	VBG	denotes	using
T25213	3344-3347	NN	denotes	RPM
T25215	3348-3353	NN	denotes	Yeast
T25216	3354-3361	NN	denotes	Plasmid
T25217	3362-3371	NN	denotes	Isolation
T25214	3372-3375	NN	denotes	Kit
T25218	3376-3377	-LRB-	denotes	(
T25219	3377-3385	NNP	denotes	Qbiogene
T25220	3385-3386	-RRB-	denotes	)
T25221	3386-3387	.	denotes	.
T25222	3387-3562	sentence	denotes	In order to confirm the protein interactions, single colonies were picked and grown individually in synthetic complete media lacking leucine, tryptophan and containing X-gal.
T25223	3388-3390	IN	denotes	In
T25225	3391-3396	NN	denotes	order
T25226	3397-3399	TO	denotes	to
T25227	3400-3407	VB	denotes	confirm
T25228	3408-3411	DT	denotes	the
T25230	3412-3419	NN	denotes	protein
T25229	3420-3432	NNS	denotes	interactions
T25231	3432-3434	,	denotes	,
T25232	3434-3440	JJ	denotes	single
T25233	3441-3449	NNS	denotes	colonies
T25234	3450-3454	VBD	denotes	were
T25224	3455-3461	VBN	denotes	picked
T25235	3462-3465	CC	denotes	and
T25236	3466-3471	VBN	denotes	grown
T25237	3472-3484	RB	denotes	individually
T25238	3485-3487	IN	denotes	in
T25239	3488-3497	JJ	denotes	synthetic
T25241	3498-3506	JJ	denotes	complete
T25240	3507-3512	NNS	denotes	media
T25242	3513-3520	VBG	denotes	lacking
T25243	3521-3528	NN	denotes	leucine
T25245	3528-3530	,	denotes	,
T25244	3530-3540	NN	denotes	tryptophan
T25246	3541-3544	CC	denotes	and
T25247	3545-3555	VBG	denotes	containing
T25248	3556-3557	NN	denotes	X
T25250	3557-3558	HYPH	denotes	-
T25249	3558-3561	NN	denotes	gal
T25251	3561-3562	.	denotes	.
T25252	3562-3628	sentence	denotes	After 4–5 days, X-gal positive clones were selected and analyzed.
T25253	3563-3568	IN	denotes	After
T25255	3569-3570	CD	denotes	4
T25257	3570-3571	SYM	denotes	–
T25256	3571-3572	CD	denotes	5
T25258	3573-3577	NNS	denotes	days
T25259	3577-3579	,	denotes	,
T25260	3579-3580	NN	denotes	X
T25262	3580-3581	NN	denotes	-
T25261	3581-3584	NN	denotes	gal
T25263	3585-3593	JJ	denotes	positive
T25264	3594-3600	NNS	denotes	clones
T25265	3601-3605	VBD	denotes	were
T25254	3606-3614	VBN	denotes	selected
T25266	3615-3618	CC	denotes	and
T25267	3619-3627	VBN	denotes	analyzed
T25268	3627-3628	.	denotes	.
T25269	3628-3800	sentence	denotes	To test self-interaction of mr-s, we subcloned the full-length, N-terminus (amino acids 1–400) and C-terminus (amino acids 391–542) of mr-s into pGBKT7 and pGADT7 vectors.
T25270	3629-3631	TO	denotes	To
T25271	3632-3636	VB	denotes	test
T25273	3637-3641	NN	denotes	self
T25275	3641-3642	HYPH	denotes	-
T25274	3642-3653	NN	denotes	interaction
T25276	3654-3656	IN	denotes	of
T25277	3657-3659	NN	denotes	mr
T25279	3659-3660	HYPH	denotes	-
T25278	3660-3661	NN	denotes	s
T25280	3661-3663	,	denotes	,
T25281	3663-3665	PRP	denotes	we
T25272	3666-3675	VBD	denotes	subcloned
T25282	3676-3679	DT	denotes	the
T25284	3680-3684	JJ	denotes	full
T25285	3684-3685	HYPH	denotes	-
T25283	3685-3691	NN	denotes	length
T25286	3691-3693	,	denotes	,
T25287	3693-3694	NN	denotes	N
T25289	3694-3695	HYPH	denotes	-
T25288	3695-3703	NN	denotes	terminus
T25290	3704-3705	-LRB-	denotes	(
T25292	3705-3710	NN	denotes	amino
T25293	3711-3716	NNS	denotes	acids
T25291	3717-3718	CD	denotes	1
T25294	3718-3719	SYM	denotes	–
T25295	3719-3722	CD	denotes	400
T25296	3722-3723	-RRB-	denotes	)
T25297	3724-3727	CC	denotes	and
T25298	3728-3729	NN	denotes	C
T25300	3729-3730	HYPH	denotes	-
T25299	3730-3738	NN	denotes	terminus
T25301	3739-3740	-LRB-	denotes	(
T25303	3740-3745	NN	denotes	amino
T25304	3746-3751	NNS	denotes	acids
T25302	3752-3755	CD	denotes	391
T25305	3755-3756	SYM	denotes	–
T25306	3756-3759	CD	denotes	542
T25307	3759-3760	-RRB-	denotes	)
T25308	3761-3763	IN	denotes	of
T25309	3764-3766	NN	denotes	mr
T25311	3766-3767	HYPH	denotes	-
T25310	3767-3768	NN	denotes	s
T25312	3769-3773	IN	denotes	into
T25313	3774-3780	NN	denotes	pGBKT7
T25315	3781-3784	CC	denotes	and
T25316	3785-3791	NN	denotes	pGADT7
T25314	3792-3799	NNS	denotes	vectors
T25317	3799-3800	.	denotes	.
T25318	3800-3885	sentence	denotes	We assessed interactions by scoring blue color on plates of medium containing X-gal.
T25319	3801-3803	PRP	denotes	We
T25320	3804-3812	VBD	denotes	assessed
T25321	3813-3825	NNS	denotes	interactions
T25322	3826-3828	IN	denotes	by
T25323	3829-3836	VBG	denotes	scoring
T25324	3837-3841	JJ	denotes	blue
T25325	3842-3847	NN	denotes	color
T25326	3848-3850	IN	denotes	on
T25327	3851-3857	NNS	denotes	plates
T25328	3858-3860	IN	denotes	of
T25329	3861-3867	NN	denotes	medium
T25330	3868-3878	VBG	denotes	containing
T25331	3879-3880	NN	denotes	X
T25333	3880-3881	HYPH	denotes	-
T25332	3881-3884	NN	denotes	gal
T25334	3884-3885	.	denotes	.
T26198	3887-3906	NN	denotes	Immunoprecipitation
T26199	3907-3912	NN	denotes	assay
T26200	3912-4238	sentence	denotes	For immunoprecipitation assay, 4× hemagglutinin (HA) or 3× Flag tagged cDNA fragment encoding full-length mr-s (full-HA and Flag-mrs), a cDNA fragment encoding amino acids 1–400 (ΔSAM-HA and Flag-ΔSAM), and a cDNA fragment encoding amino acids 400–542 (Flag-SAM) were subcloned into the pcDNA3 (Invitrogen) expression vector.
T26201	3913-3916	IN	denotes	For
T26203	3917-3936	NN	denotes	immunoprecipitation
T26204	3937-3942	NN	denotes	assay
T26205	3942-3944	,	denotes	,
T26206	3944-3945	CD	denotes	4
T26208	3945-3946	SYM	denotes	×
T26207	3947-3960	NN	denotes	hemagglutinin
T26210	3961-3962	-LRB-	denotes	(
T26211	3962-3964	NN	denotes	HA
T26212	3964-3965	-RRB-	denotes	)
T26213	3966-3968	CC	denotes	or
T26214	3969-3970	CD	denotes	3
T26216	3970-3971	SYM	denotes	×
T26215	3972-3976	NN	denotes	Flag
T26209	3977-3983	VBN	denotes	tagged
T26218	3984-3988	NN	denotes	cDNA
T26217	3989-3997	NN	denotes	fragment
T26219	3998-4006	VBG	denotes	encoding
T26220	4007-4011	JJ	denotes	full
T26222	4011-4012	HYPH	denotes	-
T26221	4012-4018	NN	denotes	length
T26224	4019-4021	NN	denotes	mr
T26225	4021-4022	HYPH	denotes	-
T26223	4022-4023	NN	denotes	s
T26226	4024-4025	-LRB-	denotes	(
T26228	4025-4029	JJ	denotes	full
T26229	4029-4030	HYPH	denotes	-
T26227	4030-4032	NN	denotes	HA
T26230	4033-4036	CC	denotes	and
T26231	4037-4041	NN	denotes	Flag
T26233	4041-4042	HYPH	denotes	-
T26232	4042-4045	NN	denotes	mrs
T26234	4045-4046	-RRB-	denotes	)
T26235	4046-4048	,	denotes	,
T26236	4048-4049	DT	denotes	a
T26238	4050-4054	NN	denotes	cDNA
T26237	4055-4063	NN	denotes	fragment
T26239	4064-4072	VBG	denotes	encoding
T26240	4073-4078	NN	denotes	amino
T26242	4079-4084	NNS	denotes	acids
T26241	4085-4086	CD	denotes	1
T26243	4086-4087	SYM	denotes	–
T26244	4087-4090	CD	denotes	400
T26245	4091-4092	-LRB-	denotes	(
T26247	4092-4096	NN	denotes	ΔSAM
T26248	4096-4097	HYPH	denotes	-
T26246	4097-4099	NN	denotes	HA
T26249	4100-4103	CC	denotes	and
T26250	4104-4108	NN	denotes	Flag
T26252	4108-4109	HYPH	denotes	-
T26251	4109-4113	NN	denotes	ΔSAM
T26253	4113-4114	-RRB-	denotes	)
T26254	4114-4116	,	denotes	,
T26255	4116-4119	CC	denotes	and
T26256	4120-4121	DT	denotes	a
T26258	4122-4126	NN	denotes	cDNA
T26257	4127-4135	NN	denotes	fragment
T26259	4136-4144	VBG	denotes	encoding
T26260	4145-4150	NN	denotes	amino
T26262	4151-4156	NNS	denotes	acids
T26261	4157-4160	CD	denotes	400
T26263	4160-4161	SYM	denotes	–
T26264	4161-4164	CD	denotes	542
T26265	4165-4166	-LRB-	denotes	(
T26267	4166-4170	NN	denotes	Flag
T26268	4170-4171	HYPH	denotes	-
T26266	4171-4174	NN	denotes	SAM
T26269	4174-4175	-RRB-	denotes	)
T26270	4176-4180	VBD	denotes	were
T26202	4181-4190	VBN	denotes	subcloned
T26271	4191-4195	IN	denotes	into
T26272	4196-4199	DT	denotes	the
T26274	4200-4206	NN	denotes	pcDNA3
T26275	4207-4208	-LRB-	denotes	(
T26276	4208-4218	NNP	denotes	Invitrogen
T26277	4218-4219	-RRB-	denotes	)
T26278	4220-4230	NN	denotes	expression
T26273	4231-4237	NN	denotes	vector
T26279	4237-4238	.	denotes	.
T26280	4238-4323	sentence	denotes	We also constructed two site-directed mutants, Flag-W404A and Flag-G453A, using PCR.
T26281	4239-4241	PRP	denotes	We
T26283	4242-4246	RB	denotes	also
T26282	4247-4258	VBD	denotes	constructed
T26284	4259-4262	CD	denotes	two
T26286	4263-4267	NN	denotes	site
T26288	4267-4268	HYPH	denotes	-
T26287	4268-4276	VBN	denotes	directed
T26285	4277-4284	NNS	denotes	mutants
T26289	4284-4286	,	denotes	,
T26290	4286-4290	NN	denotes	Flag
T26292	4290-4291	HYPH	denotes	-
T26291	4291-4296	NN	denotes	W404A
T26293	4297-4300	CC	denotes	and
T26294	4301-4305	NN	denotes	Flag
T26296	4305-4306	HYPH	denotes	-
T26295	4306-4311	NN	denotes	G453A
T26297	4311-4313	,	denotes	,
T26298	4313-4318	VBG	denotes	using
T26299	4319-4322	NN	denotes	PCR
T26300	4322-4323	.	denotes	.
T26301	4323-4383	sentence	denotes	Each of these mutations was also introduced into Flag-mr-s.
T26302	4324-4328	DT	denotes	Each
T26304	4329-4331	IN	denotes	of
T26305	4332-4337	DT	denotes	these
T26306	4338-4347	NNS	denotes	mutations
T26307	4348-4351	VBD	denotes	was
T26308	4352-4356	RB	denotes	also
T26303	4357-4367	VBN	denotes	introduced
T26309	4368-4372	IN	denotes	into
T26310	4373-4377	NN	denotes	Flag
T26311	4377-4378	HYPH	denotes	-
T26312	4378-4380	NN	denotes	mr
T26314	4380-4381	HYPH	denotes	-
T26313	4381-4382	NN	denotes	s
T26315	4382-4383	.	denotes	.
T26316	4383-4480	sentence	denotes	We transfected HEK293T cells with 5 μg of plasmid DNA per 6 cm dish by calcium phosphate method.
T26317	4384-4386	PRP	denotes	We
T26318	4387-4398	VBD	denotes	transfected
T26319	4399-4406	NN	denotes	HEK293T
T26320	4407-4412	NNS	denotes	cells
T26321	4413-4417	IN	denotes	with
T26322	4418-4419	CD	denotes	5
T26323	4420-4422	NN	denotes	μg
T26324	4423-4425	IN	denotes	of
T26325	4426-4433	NN	denotes	plasmid
T26326	4434-4437	NN	denotes	DNA
T26327	4438-4441	IN	denotes	per
T26328	4442-4443	CD	denotes	6
T26329	4444-4446	NN	denotes	cm
T26330	4447-4451	NN	denotes	dish
T26331	4452-4454	IN	denotes	by
T26332	4455-4462	NN	denotes	calcium
T26333	4463-4472	NN	denotes	phosphate
T26334	4473-4479	NN	denotes	method
T26335	4479-4480	.	denotes	.
T26336	4480-4771	sentence	denotes	Approximately 48 hr after transfection, cells were harvested in immunoprecipitation buffer (50 mM Tris-HCl [pH 7.5], 1 mM EDTA, 2 mM MgCl2, 150 mM NaCl, 1 mM phenylmethylsulfonyl fluoride [Wako], 1% Nonidet P-40, 10% glycerol) in the presence of protease inhibitor cocktail tablets (Roche).
T26337	4481-4494	RB	denotes	Approximately
T26338	4495-4497	CD	denotes	48
T26339	4498-4500	NN	denotes	hr
T26340	4501-4506	IN	denotes	after
T26342	4507-4519	NN	denotes	transfection
T26343	4519-4521	,	denotes	,
T26344	4521-4526	NNS	denotes	cells
T26345	4527-4531	VBD	denotes	were
T26341	4532-4541	VBN	denotes	harvested
T26346	4542-4544	IN	denotes	in
T26347	4545-4564	NN	denotes	immunoprecipitation
T26348	4565-4571	NN	denotes	buffer
T26349	4572-4573	-LRB-	denotes	(
T26351	4573-4575	CD	denotes	50
T26352	4576-4578	NN	denotes	mM
T26353	4579-4583	NN	denotes	Tris
T26354	4583-4584	HYPH	denotes	-
T26350	4584-4587	NN	denotes	HCl
T26355	4588-4589	-LRB-	denotes	[
T26356	4589-4591	NN	denotes	pH
T26357	4592-4595	CD	denotes	7.5
T26358	4595-4596	-RRB-	denotes	]
T26359	4596-4598	,	denotes	,
T26360	4598-4599	CD	denotes	1
T26361	4600-4602	NN	denotes	mM
T26362	4603-4607	NN	denotes	EDTA
T26363	4607-4609	,	denotes	,
T26364	4609-4610	CD	denotes	2
T26365	4611-4613	NN	denotes	mM
T26366	4614-4619	NN	denotes	MgCl2
T26367	4619-4621	,	denotes	,
T26368	4621-4624	CD	denotes	150
T26369	4625-4627	NN	denotes	mM
T26370	4628-4632	NN	denotes	NaCl
T26371	4632-4634	,	denotes	,
T26372	4634-4635	CD	denotes	1
T26373	4636-4638	NN	denotes	mM
T26375	4639-4659	NN	denotes	phenylmethylsulfonyl
T26374	4660-4668	NN	denotes	fluoride
T26376	4669-4670	-LRB-	denotes	[
T26377	4670-4674	NN	denotes	Wako
T26378	4674-4675	-RRB-	denotes	]
T26379	4675-4677	,	denotes	,
T26380	4677-4678	CD	denotes	1
T26381	4678-4679	NN	denotes	%
T26383	4680-4687	NN	denotes	Nonidet
T26382	4688-4689	NN	denotes	P
T26384	4689-4690	HYPH	denotes	-
T26385	4690-4692	CD	denotes	40
T26386	4692-4694	,	denotes	,
T26387	4694-4696	CD	denotes	10
T26388	4696-4697	NN	denotes	%
T26389	4698-4706	NN	denotes	glycerol
T26390	4706-4707	-RRB-	denotes	)
T26391	4708-4710	IN	denotes	in
T26392	4711-4714	DT	denotes	the
T26393	4715-4723	NN	denotes	presence
T26394	4724-4726	IN	denotes	of
T26395	4727-4735	NN	denotes	protease
T26396	4736-4745	NN	denotes	inhibitor
T26398	4746-4754	NN	denotes	cocktail
T26397	4755-4762	NNS	denotes	tablets
T26399	4763-4764	-LRB-	denotes	(
T26400	4764-4769	NNP	denotes	Roche
T26401	4769-4770	-RRB-	denotes	)
T26402	4770-4771	.	denotes	.
T26403	4771-4951	sentence	denotes	For each reaction, 1 mg of cell lysate was mixed with 0.5 μg of anti-Flag antibody (SIGMA, F3165) and 15 μl of protein G-Sepharose (Amersham) on a rotating wheel at 4°C for 2 hrs.
T26404	4772-4775	IN	denotes	For
T26406	4776-4780	DT	denotes	each
T26407	4781-4789	NN	denotes	reaction
T26408	4789-4791	,	denotes	,
T26409	4791-4792	CD	denotes	1
T26410	4793-4795	NN	denotes	mg
T26411	4796-4798	IN	denotes	of
T26412	4799-4803	NN	denotes	cell
T26413	4804-4810	NN	denotes	lysate
T26414	4811-4814	VBD	denotes	was
T26405	4815-4820	VBN	denotes	mixed
T26415	4821-4825	IN	denotes	with
T26416	4826-4829	CD	denotes	0.5
T26417	4830-4832	NN	denotes	μg
T26418	4833-4835	IN	denotes	of
T26419	4836-4845	JJ	denotes	anti-Flag
T26420	4846-4854	NN	denotes	antibody
T26421	4855-4856	-LRB-	denotes	(
T26423	4856-4861	NNP	denotes	SIGMA
T26424	4861-4863	,	denotes	,
T26422	4863-4868	NN	denotes	F3165
T26425	4868-4869	-RRB-	denotes	)
T26426	4870-4873	CC	denotes	and
T26427	4874-4876	CD	denotes	15
T26428	4877-4879	NN	denotes	μl
T26429	4880-4882	IN	denotes	of
T26430	4883-4890	NN	denotes	protein
T26432	4891-4892	NN	denotes	G
T26433	4892-4893	HYPH	denotes	-
T26431	4893-4902	NN	denotes	Sepharose
T26434	4903-4904	-LRB-	denotes	(
T26435	4904-4912	NNP	denotes	Amersham
T26436	4912-4913	-RRB-	denotes	)
T26437	4914-4916	IN	denotes	on
T26438	4917-4918	DT	denotes	a
T26440	4919-4927	VBG	denotes	rotating
T26439	4928-4933	NN	denotes	wheel
T26441	4934-4936	IN	denotes	at
T26442	4937-4938	CD	denotes	4
T26443	4938-4940	NN	denotes	°C
T26444	4941-4944	IN	denotes	for
T26445	4945-4946	CD	denotes	2
T26446	4947-4950	NNS	denotes	hrs
T26447	4950-4951	.	denotes	.
T26448	4951-5030	sentence	denotes	Protein concentration was determined by the BCA protein assay system (Pierce).
T26449	4952-4959	NN	denotes	Protein
T26450	4960-4973	NN	denotes	concentration
T26452	4974-4977	VBD	denotes	was
T26451	4978-4988	VBN	denotes	determined
T26453	4989-4991	IN	denotes	by
T26454	4992-4995	DT	denotes	the
T26456	4996-4999	NN	denotes	BCA
T26458	5000-5007	NN	denotes	protein
T26457	5008-5013	NN	denotes	assay
T26455	5014-5020	NN	denotes	system
T26459	5021-5022	-LRB-	denotes	(
T26460	5022-5028	NNP	denotes	Pierce
T26461	5028-5029	-RRB-	denotes	)
T26462	5029-5030	.	denotes	.
T26463	5030-5199	sentence	denotes	The beads were then washed three times with immunoprecipitation buffer and followed by three times with wash buffer (50 mM Tris-HCl [pH 7.5], 150 mM NaCl, 20 mM MgCl2).
T26464	5031-5034	DT	denotes	The
T26465	5035-5040	NNS	denotes	beads
T26467	5041-5045	VBD	denotes	were
T26468	5046-5050	RB	denotes	then
T26466	5051-5057	VBN	denotes	washed
T26469	5058-5063	CD	denotes	three
T26470	5064-5069	NNS	denotes	times
T26471	5070-5074	IN	denotes	with
T26472	5075-5094	NN	denotes	immunoprecipitation
T26473	5095-5101	NN	denotes	buffer
T26474	5102-5105	CC	denotes	and
T26475	5106-5114	VBN	denotes	followed
T26476	5115-5117	IN	denotes	by
T26477	5118-5123	CD	denotes	three
T26478	5124-5129	NNS	denotes	times
T26479	5130-5134	IN	denotes	with
T26480	5135-5139	NN	denotes	wash
T26481	5140-5146	NN	denotes	buffer
T26482	5147-5148	-LRB-	denotes	(
T26484	5148-5150	CD	denotes	50
T26485	5151-5153	NN	denotes	mM
T26486	5154-5158	NN	denotes	Tris
T26487	5158-5159	HYPH	denotes	-
T26483	5159-5162	NN	denotes	HCl
T26488	5163-5164	-LRB-	denotes	[
T26489	5164-5166	NN	denotes	pH
T26490	5167-5170	CD	denotes	7.5
T26491	5170-5171	-RRB-	denotes	]
T26492	5171-5173	,	denotes	,
T26493	5173-5176	CD	denotes	150
T26494	5177-5179	NN	denotes	mM
T26495	5180-5184	NN	denotes	NaCl
T26496	5184-5186	,	denotes	,
T26497	5186-5188	CD	denotes	20
T26498	5189-5191	NN	denotes	mM
T26499	5192-5197	NN	denotes	MgCl2
T26500	5197-5198	-RRB-	denotes	)
T26501	5198-5199	.	denotes	.
T26502	5199-5328	sentence	denotes	Proteins were then boiled for 5 min in SDS sample buffer (1% SDS, 1 mM Tris [pH 6.8], 40 mM DTT, 4% glycerol, 0.01% pyronine Y).
T26503	5200-5208	NNS	denotes	Proteins
T26505	5209-5213	VBD	denotes	were
T26506	5214-5218	RB	denotes	then
T26504	5219-5225	VBN	denotes	boiled
T26507	5226-5229	IN	denotes	for
T26508	5230-5231	CD	denotes	5
T26509	5232-5235	NN	denotes	min
T26510	5236-5238	IN	denotes	in
T26511	5239-5242	NN	denotes	SDS
T26513	5243-5249	NN	denotes	sample
T26512	5250-5256	NN	denotes	buffer
T26514	5257-5258	-LRB-	denotes	(
T26516	5258-5259	CD	denotes	1
T26517	5259-5260	NN	denotes	%
T26515	5261-5264	NN	denotes	SDS
T26518	5264-5266	,	denotes	,
T26519	5266-5267	CD	denotes	1
T26520	5268-5270	NN	denotes	mM
T26521	5271-5275	NN	denotes	Tris
T26522	5276-5277	-LRB-	denotes	[
T26523	5277-5279	NN	denotes	pH
T26524	5280-5283	CD	denotes	6.8
T26525	5283-5284	-RRB-	denotes	]
T26526	5284-5286	,	denotes	,
T26527	5286-5288	CD	denotes	40
T26528	5289-5291	NN	denotes	mM
T26529	5292-5295	NN	denotes	DTT
T26530	5295-5297	,	denotes	,
T26531	5297-5298	CD	denotes	4
T26532	5298-5299	NN	denotes	%
T26533	5300-5308	NN	denotes	glycerol
T26534	5308-5310	,	denotes	,
T26535	5310-5314	CD	denotes	0.01
T26536	5314-5315	NN	denotes	%
T26538	5316-5324	NN	denotes	pyronine
T26537	5325-5326	NN	denotes	Y
T26539	5326-5327	-RRB-	denotes	)
T26540	5327-5328	.	denotes	.
T26541	5328-5521	sentence	denotes	The supernatants were fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to the nitrocellulose membrane (Trans-Blot Transfer Medium, Bio-Rad).
T26542	5329-5332	DT	denotes	The
T26543	5333-5345	NNS	denotes	supernatants
T26545	5346-5350	VBD	denotes	were
T26544	5351-5363	VBN	denotes	fractionated
T26546	5364-5366	IN	denotes	by
T26547	5367-5373	NN	denotes	sodium
T26549	5374-5381	NN	denotes	dodecyl
T26548	5382-5389	NN	denotes	sulfate
T26551	5389-5390	HYPH	denotes	-
T26552	5390-5404	NN	denotes	polyacrylamide
T26550	5405-5408	NN	denotes	gel
T26553	5409-5424	NN	denotes	electrophoresis
T26554	5425-5426	-LRB-	denotes	(
T26556	5426-5429	NN	denotes	SDS
T26557	5429-5430	HYPH	denotes	-
T26555	5430-5434	NN	denotes	PAGE
T26558	5434-5435	-RRB-	denotes	)
T26559	5436-5439	CC	denotes	and
T26560	5440-5451	VBN	denotes	transferred
T26561	5452-5454	IN	denotes	to
T26562	5455-5458	DT	denotes	the
T26564	5459-5473	NN	denotes	nitrocellulose
T26563	5474-5482	NN	denotes	membrane
T26565	5483-5484	-LRB-	denotes	(
T26567	5484-5489	NN	denotes	Trans
T26569	5489-5490	HYPH	denotes	-
T26568	5490-5494	NN	denotes	Blot
T26570	5495-5503	NN	denotes	Transfer
T26566	5504-5510	NN	denotes	Medium
T26571	5510-5512	,	denotes	,
T26572	5512-5515	NNP	denotes	Bio
T26574	5515-5516	HYPH	denotes	-
T26573	5516-5519	NNP	denotes	Rad
T26575	5519-5520	-RRB-	denotes	)
T26576	5520-5521	.	denotes	.
T26577	5521-5615	sentence	denotes	Western blotting was performed with rabbit polyclonal anti-HA antibody (Santa Cruz, #sc-805).
T26578	5522-5529	NNP	denotes	Western
T26579	5530-5538	NN	denotes	blotting
T26581	5539-5542	VBD	denotes	was
T26580	5543-5552	VBN	denotes	performed
T26582	5553-5557	IN	denotes	with
T26583	5558-5564	NN	denotes	rabbit
T26585	5565-5575	JJ	denotes	polyclonal
T26586	5576-5583	JJ	denotes	anti-HA
T26584	5584-5592	NN	denotes	antibody
T26587	5593-5594	-LRB-	denotes	(
T26589	5594-5599	NNP	denotes	Santa
T26588	5600-5604	NNP	denotes	Cruz
T26590	5604-5606	,	denotes	,
T26591	5606-5607	SYM	denotes	#
T26592	5607-5609	NN	denotes	sc
T26593	5609-5610	HYPH	denotes	-
T26594	5610-5613	CD	denotes	805
T26595	5613-5614	-RRB-	denotes	)
T26596	5614-5615	.	denotes	.
T26597	5615-5757	sentence	denotes	Signals were detected with horseradish peroxidase- conjugated goat anti-rabbit IgG and ECL plus Western Blotting Detection System (Amersham).
T26598	5616-5623	NNS	denotes	Signals
T26600	5624-5628	VBD	denotes	were
T26599	5629-5637	VBN	denotes	detected
T26601	5638-5642	IN	denotes	with
T26602	5643-5654	NN	denotes	horseradish
T26603	5655-5665	NN	denotes	peroxidase
T26605	5665-5666	HYPH	denotes	-
T26604	5667-5677	VBN	denotes	conjugated
T26607	5678-5682	NN	denotes	goat
T26608	5683-5694	JJ	denotes	anti-rabbit
T26606	5695-5698	NN	denotes	IgG
T26609	5699-5702	CC	denotes	and
T26610	5703-5706	NN	denotes	ECL
T26612	5707-5711	CC	denotes	plus
T26613	5712-5719	NNP	denotes	Western
T26614	5720-5728	NN	denotes	Blotting
T26615	5729-5738	NN	denotes	Detection
T26611	5739-5745	NN	denotes	System
T26616	5746-5747	-LRB-	denotes	(
T26617	5747-5755	NNP	denotes	Amersham
T26618	5755-5756	-RRB-	denotes	)
T26619	5756-5757	.	denotes	.
T27028	5759-5770	JJ	denotes	Subcellular
T27030	5771-5783	NN	denotes	localization
T27029	5784-5792	NN	denotes	analysis
T27031	5792-6068	sentence	denotes	We transfected the plasmid encoding HA-tagged full-length mr-s into HEK293T cells, seeded cells on coverslips coated with collagen 48 hr after transfection, fixed with 4% paraformaldehyde in phosphate-buffered saline (PBS) at room temperature for 20 min, and washed with PBS.
T27032	5793-5795	PRP	denotes	We
T27033	5796-5807	VBD	denotes	transfected
T27034	5808-5811	DT	denotes	the
T27035	5812-5819	NN	denotes	plasmid
T27036	5820-5828	VBG	denotes	encoding
T27037	5829-5831	NN	denotes	HA
T27039	5831-5832	HYPH	denotes	-
T27038	5832-5838	VBN	denotes	tagged
T27041	5839-5843	JJ	denotes	full
T27043	5843-5844	HYPH	denotes	-
T27042	5844-5850	NN	denotes	length
T27044	5851-5853	NN	denotes	mr
T27045	5853-5854	HYPH	denotes	-
T27040	5854-5855	NN	denotes	s
T27046	5856-5860	IN	denotes	into
T27047	5861-5868	NN	denotes	HEK293T
T27048	5869-5874	NNS	denotes	cells
T27049	5874-5876	,	denotes	,
T27050	5876-5882	VBD	denotes	seeded
T27051	5883-5888	NNS	denotes	cells
T27052	5889-5891	IN	denotes	on
T27053	5892-5902	NNS	denotes	coverslips
T27054	5903-5909	VBN	denotes	coated
T27055	5910-5914	IN	denotes	with
T27056	5915-5923	NN	denotes	collagen
T27057	5924-5926	CD	denotes	48
T27058	5927-5929	NN	denotes	hr
T27059	5930-5935	IN	denotes	after
T27060	5936-5948	NN	denotes	transfection
T27061	5948-5950	,	denotes	,
T27062	5950-5955	VBN	denotes	fixed
T27063	5956-5960	IN	denotes	with
T27064	5961-5962	CD	denotes	4
T27065	5962-5963	NN	denotes	%
T27066	5964-5980	NN	denotes	paraformaldehyde
T27067	5981-5983	IN	denotes	in
T27068	5984-5993	NN	denotes	phosphate
T27070	5993-5994	HYPH	denotes	-
T27069	5994-6002	VBN	denotes	buffered
T27071	6003-6009	NN	denotes	saline
T27072	6010-6011	-LRB-	denotes	(
T27073	6011-6014	NN	denotes	PBS
T27074	6014-6015	-RRB-	denotes	)
T27075	6016-6018	IN	denotes	at
T27076	6019-6023	NN	denotes	room
T27077	6024-6035	NN	denotes	temperature
T27078	6036-6039	IN	denotes	for
T27079	6040-6042	CD	denotes	20
T27080	6043-6046	NN	denotes	min
T27081	6046-6048	,	denotes	,
T27082	6048-6051	CC	denotes	and
T27083	6052-6058	VBN	denotes	washed
T27084	6059-6063	IN	denotes	with
T27085	6064-6067	NN	denotes	PBS
T27086	6067-6068	.	denotes	.
T27087	6068-6263	sentence	denotes	Then we permeabilized cells in PBS containing 0.1% Triton X-100 for 15 min, washed again with PBS, and incubated in PBS containing 0.1% bovine serum albumin (BSA) for 30 min at room temperature.
T27088	6069-6073	RB	denotes	Then
T27090	6074-6076	PRP	denotes	we
T27089	6077-6090	VBD	denotes	permeabilized
T27091	6091-6096	NNS	denotes	cells
T27092	6097-6099	IN	denotes	in
T27093	6100-6103	NN	denotes	PBS
T27094	6104-6114	VBG	denotes	containing
T27095	6115-6118	CD	denotes	0.1
T27096	6118-6119	NN	denotes	%
T27098	6120-6126	NN	denotes	Triton
T27099	6127-6128	NN	denotes	X
T27100	6128-6129	HYPH	denotes	-
T27097	6129-6132	NN	denotes	100
T27101	6133-6136	IN	denotes	for
T27102	6137-6139	CD	denotes	15
T27103	6140-6143	NN	denotes	min
T27104	6143-6145	,	denotes	,
T27105	6145-6151	VBD	denotes	washed
T27106	6152-6157	RB	denotes	again
T27107	6158-6162	IN	denotes	with
T27108	6163-6166	NN	denotes	PBS
T27109	6166-6168	,	denotes	,
T27110	6168-6171	CC	denotes	and
T27111	6172-6181	VBD	denotes	incubated
T27112	6182-6184	IN	denotes	in
T27113	6185-6188	NN	denotes	PBS
T27114	6189-6199	VBG	denotes	containing
T27115	6200-6203	CD	denotes	0.1
T27116	6203-6204	NN	denotes	%
T27117	6205-6211	JJ	denotes	bovine
T27119	6212-6217	NN	denotes	serum
T27118	6218-6225	NN	denotes	albumin
T27120	6226-6227	-LRB-	denotes	(
T27121	6227-6230	NN	denotes	BSA
T27122	6230-6231	-RRB-	denotes	)
T27123	6232-6235	IN	denotes	for
T27124	6236-6238	CD	denotes	30
T27125	6239-6242	NN	denotes	min
T27126	6243-6245	IN	denotes	at
T27127	6246-6250	NN	denotes	room
T27128	6251-6262	NN	denotes	temperature
T27129	6262-6263	.	denotes	.
T27130	6263-6357	sentence	denotes	We incubated cells with anti-HA antibody overnight at 4°C (1:200 in PBS containing 0.2% BSA).
T27131	6264-6266	PRP	denotes	We
T27132	6267-6276	VBD	denotes	incubated
T27133	6277-6282	NNS	denotes	cells
T27134	6283-6287	IN	denotes	with
T27135	6288-6295	JJ	denotes	anti-HA
T27136	6296-6304	NN	denotes	antibody
T27137	6305-6314	RB	denotes	overnight
T27138	6315-6317	IN	denotes	at
T27139	6318-6319	CD	denotes	4
T27140	6319-6321	NN	denotes	°C
T27141	6322-6323	-LRB-	denotes	(
T27142	6323-6324	CD	denotes	1
T27143	6324-6325	SYM	denotes	:
T27144	6325-6328	CD	denotes	200
T27145	6329-6331	IN	denotes	in
T27146	6332-6335	NN	denotes	PBS
T27147	6336-6346	VBG	denotes	containing
T27148	6347-6350	CD	denotes	0.2
T27149	6350-6351	NN	denotes	%
T27150	6352-6355	NN	denotes	BSA
T27151	6355-6356	-RRB-	denotes	)
T27152	6356-6357	.	denotes	.
T27153	6357-6540	sentence	denotes	The following day, we washed cells three times with PBS and incubated with a Cy3-conjugated goat IgG against rabbit IgG (1:400 in PBS, Jackson ImmunoReseach Laboratories) for 30 min.
T27154	6358-6361	DT	denotes	The
T27156	6362-6371	VBG	denotes	following
T27155	6372-6375	NN	denotes	day
T27158	6375-6377	,	denotes	,
T27159	6377-6379	PRP	denotes	we
T27157	6380-6386	VBD	denotes	washed
T27160	6387-6392	NNS	denotes	cells
T27161	6393-6398	CD	denotes	three
T27162	6399-6404	NNS	denotes	times
T27163	6405-6409	IN	denotes	with
T27164	6410-6413	NN	denotes	PBS
T27165	6414-6417	CC	denotes	and
T27166	6418-6427	VBD	denotes	incubated
T27167	6428-6432	IN	denotes	with
T27168	6433-6434	DT	denotes	a
T27170	6435-6438	NN	denotes	Cy3
T27172	6438-6439	HYPH	denotes	-
T27171	6439-6449	VBN	denotes	conjugated
T27173	6450-6454	NN	denotes	goat
T27169	6455-6458	NN	denotes	IgG
T27174	6459-6466	IN	denotes	against
T27175	6467-6473	NN	denotes	rabbit
T27176	6474-6477	NN	denotes	IgG
T27177	6478-6479	-LRB-	denotes	(
T27179	6479-6480	CD	denotes	1
T27180	6480-6481	SYM	denotes	:
T27181	6481-6484	CD	denotes	400
T27182	6485-6487	IN	denotes	in
T27183	6488-6491	NN	denotes	PBS
T27184	6491-6493	,	denotes	,
T27185	6493-6500	NNP	denotes	Jackson
T27186	6501-6514	NNP	denotes	ImmunoReseach
T27178	6515-6527	NNP	denotes	Laboratories
T27187	6527-6528	-RRB-	denotes	)
T27188	6529-6532	IN	denotes	for
T27189	6533-6535	CD	denotes	30
T27190	6536-6539	NN	denotes	min
T27191	6539-6540	.	denotes	.
T27192	6540-6682	sentence	denotes	We rinsed cells three times with PBS, followed by observation using a confocal microscope FV300 (Olympus) equipped with a 60× objective lens.
T27193	6541-6543	PRP	denotes	We
T27194	6544-6550	VBD	denotes	rinsed
T27195	6551-6556	NNS	denotes	cells
T27196	6557-6562	CD	denotes	three
T27197	6563-6568	NNS	denotes	times
T27198	6569-6573	IN	denotes	with
T27199	6574-6577	NN	denotes	PBS
T27200	6577-6579	,	denotes	,
T27201	6579-6587	VBN	denotes	followed
T27202	6588-6590	IN	denotes	by
T27203	6591-6602	NN	denotes	observation
T27204	6603-6608	VBG	denotes	using
T27205	6609-6610	DT	denotes	a
T27207	6611-6619	JJ	denotes	confocal
T27208	6620-6630	NN	denotes	microscope
T27206	6631-6636	NN	denotes	FV300
T27209	6637-6638	-LRB-	denotes	(
T27210	6638-6645	NNP	denotes	Olympus
T27211	6645-6646	-RRB-	denotes	)
T27212	6647-6655	VBN	denotes	equipped
T27213	6656-6660	IN	denotes	with
T27214	6661-6662	DT	denotes	a
T27216	6663-6665	CD	denotes	60
T27217	6665-6666	SYM	denotes	×
T27218	6667-6676	NN	denotes	objective
T27215	6677-6681	NN	denotes	lens
T27219	6681-6682	.	denotes	.
T27909	6684-6694	NN	denotes	Luciferase
T27910	6695-6700	NN	denotes	assay
T27911	6700-6925	sentence	denotes	For transcriptional analysis of the 1.2-kb promoter region of mr-s, we constructed a reporter plasmid (pro1.2k) by subcloning a 1.2-kb upstream genomic fragment of mr-s gene into a pGL3 luciferase reporter plasmid (Promega).
T27912	6701-6704	IN	denotes	For
T27914	6705-6720	JJ	denotes	transcriptional
T27915	6721-6729	NN	denotes	analysis
T27916	6730-6732	IN	denotes	of
T27917	6733-6736	DT	denotes	the
T27919	6737-6740	CD	denotes	1.2
T27921	6740-6741	HYPH	denotes	-
T27920	6741-6743	NN	denotes	kb
T27922	6744-6752	NN	denotes	promoter
T27918	6753-6759	NN	denotes	region
T27923	6760-6762	IN	denotes	of
T27924	6763-6765	NN	denotes	mr
T27926	6765-6766	HYPH	denotes	-
T27925	6766-6767	NN	denotes	s
T27927	6767-6769	,	denotes	,
T27928	6769-6771	PRP	denotes	we
T27913	6772-6783	VBD	denotes	constructed
T27929	6784-6785	DT	denotes	a
T27931	6786-6794	NN	denotes	reporter
T27930	6795-6802	NN	denotes	plasmid
T27932	6803-6804	-LRB-	denotes	(
T27933	6804-6811	NN	denotes	pro1.2k
T27934	6811-6812	-RRB-	denotes	)
T27935	6813-6815	IN	denotes	by
T27936	6816-6826	VBG	denotes	subcloning
T27937	6827-6828	DT	denotes	a
T27939	6829-6832	CD	denotes	1.2
T27941	6832-6833	HYPH	denotes	-
T27940	6833-6835	NN	denotes	kb
T27942	6836-6844	JJ	denotes	upstream
T27943	6845-6852	JJ	denotes	genomic
T27938	6853-6861	NN	denotes	fragment
T27944	6862-6864	IN	denotes	of
T27945	6865-6867	NN	denotes	mr
T27947	6867-6868	HYPH	denotes	-
T27946	6868-6869	NN	denotes	s
T27948	6870-6874	NN	denotes	gene
T27949	6875-6879	IN	denotes	into
T27950	6880-6881	DT	denotes	a
T27952	6882-6886	NN	denotes	pGL3
T27954	6887-6897	NN	denotes	luciferase
T27953	6898-6906	NN	denotes	reporter
T27951	6907-6914	NN	denotes	plasmid
T27955	6915-6916	-LRB-	denotes	(
T27956	6916-6923	NNP	denotes	Promega
T27957	6923-6924	-RRB-	denotes	)
T27958	6924-6925	.	denotes	.
T27959	6925-7087	sentence	denotes	The expression vectors were constructed by subcloning full-length Crx, Otx2, Nrl genes into a pMIK expression vector (a gift from Dr. K. Maruyama), respectively.
T27960	6926-6929	DT	denotes	The
T27962	6930-6940	NN	denotes	expression
T27961	6941-6948	NNS	denotes	vectors
T27964	6949-6953	VBD	denotes	were
T27963	6954-6965	VBN	denotes	constructed
T27965	6966-6968	IN	denotes	by
T27966	6969-6979	VBG	denotes	subcloning
T27967	6980-6984	JJ	denotes	full
T27969	6984-6985	HYPH	denotes	-
T27968	6985-6991	NN	denotes	length
T27971	6992-6995	NN	denotes	Crx
T27973	6995-6997	,	denotes	,
T27974	6997-7001	NN	denotes	Otx2
T27975	7001-7003	,	denotes	,
T27972	7003-7006	NN	denotes	Nrl
T27970	7007-7012	NNS	denotes	genes
T27976	7013-7017	IN	denotes	into
T27977	7018-7019	DT	denotes	a
T27979	7020-7024	NN	denotes	pMIK
T27980	7025-7035	NN	denotes	expression
T27978	7036-7042	NN	denotes	vector
T27981	7043-7044	-LRB-	denotes	(
T27983	7044-7045	DT	denotes	a
T27982	7046-7050	NN	denotes	gift
T27984	7051-7055	IN	denotes	from
T27985	7056-7059	NNP	denotes	Dr.
T27987	7060-7062	NNP	denotes	K.
T27986	7063-7071	NNP	denotes	Maruyama
T27988	7071-7072	-RRB-	denotes	)
T27989	7072-7074	,	denotes	,
T27990	7074-7086	RB	denotes	respectively
T27991	7086-7087	.	denotes	.
T27992	7087-7204	sentence	denotes	For the "mut1259" vector, the Crx binding site at the -1259 bp position was mutated by replacing GGATTA with AGATCT.
T27993	7088-7091	IN	denotes	For
T27995	7092-7095	DT	denotes	the
T27997	7096-7097	``	denotes	"
T27998	7097-7104	NN	denotes	mut1259
T27999	7104-7105	''	denotes	"
T27996	7106-7112	NN	denotes	vector
T28000	7112-7114	,	denotes	,
T28001	7114-7117	DT	denotes	the
T28003	7118-7121	NN	denotes	Crx
T28004	7122-7129	NN	denotes	binding
T28002	7130-7134	NN	denotes	site
T28005	7135-7137	IN	denotes	at
T28006	7138-7141	DT	denotes	the
T28008	7142-7143	SYM	denotes	-
T28009	7143-7147	CD	denotes	1259
T28010	7148-7150	NN	denotes	bp
T28007	7151-7159	NN	denotes	position
T28011	7160-7163	VBD	denotes	was
T27994	7164-7171	VBN	denotes	mutated
T28012	7172-7174	IN	denotes	by
T28013	7175-7184	VBG	denotes	replacing
T28014	7185-7191	NN	denotes	GGATTA
T28015	7192-7196	IN	denotes	with
T28016	7197-7203	NN	denotes	AGATCT
T28017	7203-7204	.	denotes	.
T28018	7204-7319	sentence	denotes	For the "mut198" vector, the Crx binding site at the -198 bp position was mutated by replacing TAATCC with GAATTC.
T28019	7205-7208	IN	denotes	For
T28021	7209-7212	DT	denotes	the
T28023	7213-7214	``	denotes	"
T28024	7214-7220	NN	denotes	mut198
T28025	7220-7221	''	denotes	"
T28022	7222-7228	NN	denotes	vector
T28026	7228-7230	,	denotes	,
T28027	7230-7233	DT	denotes	the
T28029	7234-7237	NN	denotes	Crx
T28030	7238-7245	NN	denotes	binding
T28028	7246-7250	NN	denotes	site
T28031	7251-7253	IN	denotes	at
T28032	7254-7257	DT	denotes	the
T28034	7258-7259	SYM	denotes	-
T28035	7259-7262	CD	denotes	198
T28036	7263-7265	NN	denotes	bp
T28033	7266-7274	NN	denotes	position
T28037	7275-7278	VBD	denotes	was
T28020	7279-7286	VBN	denotes	mutated
T28038	7287-7289	IN	denotes	by
T28039	7290-7299	VBG	denotes	replacing
T28040	7300-7306	NN	denotes	TAATCC
T28041	7307-7311	IN	denotes	with
T28042	7312-7318	NN	denotes	GAATTC
T28043	7318-7319	.	denotes	.
T28044	7319-7432	sentence	denotes	For the "mut72" vector, the Crx binding site at the -72 bp position was mutated by replacing GGATTA with GAATTC.
T28045	7320-7323	IN	denotes	For
T28047	7324-7327	DT	denotes	the
T28049	7328-7329	``	denotes	"
T28050	7329-7334	NN	denotes	mut72
T28051	7334-7335	''	denotes	"
T28048	7336-7342	NN	denotes	vector
T28052	7342-7344	,	denotes	,
T28053	7344-7347	DT	denotes	the
T28055	7348-7351	NN	denotes	Crx
T28056	7352-7359	NN	denotes	binding
T28054	7360-7364	NN	denotes	site
T28057	7365-7367	IN	denotes	at
T28058	7368-7371	DT	denotes	the
T28060	7372-7373	SYM	denotes	-
T28061	7373-7375	CD	denotes	72
T28062	7376-7378	NN	denotes	bp
T28059	7379-7387	NN	denotes	position
T28063	7388-7391	VBD	denotes	was
T28046	7392-7399	VBN	denotes	mutated
T28064	7400-7402	IN	denotes	by
T28065	7403-7412	VBG	denotes	replacing
T28066	7413-7419	NN	denotes	GGATTA
T28067	7420-7424	IN	denotes	with
T28068	7425-7431	NN	denotes	GAATTC
T28069	7431-7432	.	denotes	.
T28070	7432-7503	sentence	denotes	For the "mut all" vector, all of three Crx binding sites were mutated.
T28071	7433-7436	IN	denotes	For
T28073	7437-7440	DT	denotes	the
T28075	7441-7442	``	denotes	"
T28076	7442-7445	NN	denotes	mut
T28077	7446-7449	NN	denotes	all
T28078	7449-7450	''	denotes	"
T28074	7451-7457	NN	denotes	vector
T28079	7457-7459	,	denotes	,
T28080	7459-7462	DT	denotes	all
T28081	7463-7465	IN	denotes	of
T28082	7466-7471	CD	denotes	three
T28084	7472-7475	NN	denotes	Crx
T28085	7476-7483	NN	denotes	binding
T28083	7484-7489	NNS	denotes	sites
T28086	7490-7494	VBD	denotes	were
T28072	7495-7502	VBN	denotes	mutated
T28087	7502-7503	.	denotes	.
T28088	7503-7769	sentence	denotes	5xGAL4-pGL3 Control reporter plasmid, which contains five copies of the GAL4 DNA recognition sequence positioned immediately upstream of SV40 minimal promoter, was kindly gifted from Dr. T. Noguchi and used for analysis of the transcriptional activity of mr-s [50].
T28089	7504-7510	NN	denotes	5xGAL4
T28091	7510-7511	HYPH	denotes	-
T28090	7511-7515	NN	denotes	pGL3
T28093	7516-7523	NN	denotes	Control
T28094	7524-7532	NN	denotes	reporter
T28092	7533-7540	NN	denotes	plasmid
T28096	7540-7542	,	denotes	,
T28097	7542-7547	WDT	denotes	which
T28098	7548-7556	VBZ	denotes	contains
T28099	7557-7561	CD	denotes	five
T28100	7562-7568	NNS	denotes	copies
T28101	7569-7571	IN	denotes	of
T28102	7572-7575	DT	denotes	the
T28104	7576-7580	NN	denotes	GAL4
T28105	7581-7584	NN	denotes	DNA
T28106	7585-7596	NN	denotes	recognition
T28103	7597-7605	NN	denotes	sequence
T28107	7606-7616	VBN	denotes	positioned
T28108	7617-7628	RB	denotes	immediately
T28109	7629-7637	IN	denotes	upstream
T28110	7638-7640	IN	denotes	of
T28111	7641-7645	NN	denotes	SV40
T28113	7646-7653	JJ	denotes	minimal
T28112	7654-7662	NN	denotes	promoter
T28114	7662-7664	,	denotes	,
T28115	7664-7667	VBD	denotes	was
T28116	7668-7674	RB	denotes	kindly
T28095	7675-7681	VBN	denotes	gifted
T28117	7682-7686	IN	denotes	from
T28118	7687-7690	NNP	denotes	Dr.
T28120	7691-7693	NNP	denotes	T.
T28119	7694-7701	NNP	denotes	Noguchi
T28121	7702-7705	CC	denotes	and
T28122	7706-7710	VBN	denotes	used
T28123	7711-7714	IN	denotes	for
T28124	7715-7723	NN	denotes	analysis
T28125	7724-7726	IN	denotes	of
T28126	7727-7730	DT	denotes	the
T28128	7731-7746	JJ	denotes	transcriptional
T28127	7747-7755	NN	denotes	activity
T28129	7756-7758	IN	denotes	of
T28130	7759-7761	NN	denotes	mr
T28132	7761-7762	HYPH	denotes	-
T28131	7762-7763	NN	denotes	s
T28133	7764-7765	-LRB-	denotes	[
T28134	7765-7767	CD	denotes	50
T28135	7767-7768	-RRB-	denotes	]
T28136	7768-7769	.	denotes	.
T28137	7769-7905	sentence	denotes	To generate effector plasmids, various deletion fragments were produced by PCR reaction and subcloned into pGBKT7 vector, respectively.
T28138	7770-7772	TO	denotes	To
T28139	7773-7781	VB	denotes	generate
T28141	7782-7790	NN	denotes	effector
T28142	7791-7799	NNS	denotes	plasmids
T28143	7799-7801	,	denotes	,
T28144	7801-7808	JJ	denotes	various
T28146	7809-7817	NN	denotes	deletion
T28145	7818-7827	NNS	denotes	fragments
T28147	7828-7832	VBD	denotes	were
T28140	7833-7841	VBN	denotes	produced
T28148	7842-7844	IN	denotes	by
T28149	7845-7848	NN	denotes	PCR
T28150	7849-7857	NN	denotes	reaction
T28151	7858-7861	CC	denotes	and
T28152	7862-7871	VBN	denotes	subcloned
T28153	7872-7876	IN	denotes	into
T28154	7877-7883	NN	denotes	pGBKT7
T28155	7884-7890	NN	denotes	vector
T28156	7890-7892	,	denotes	,
T28157	7892-7904	RB	denotes	respectively
T28158	7904-7905	.	denotes	.
T28159	7905-8019	sentence	denotes	These fragments were digested with the sequence, which encodes GAL4 DNA binding domain, and inserted into pcDNA3.
T28160	7906-7911	DT	denotes	These
T28161	7912-7921	NNS	denotes	fragments
T28163	7922-7926	VBD	denotes	were
T28162	7927-7935	VBN	denotes	digested
T28164	7936-7940	IN	denotes	with
T28165	7941-7944	DT	denotes	the
T28166	7945-7953	NN	denotes	sequence
T28167	7953-7955	,	denotes	,
T28168	7955-7960	WDT	denotes	which
T28169	7961-7968	VBZ	denotes	encodes
T28170	7969-7973	NN	denotes	GAL4
T28172	7974-7977	NN	denotes	DNA
T28173	7978-7985	NN	denotes	binding
T28171	7986-7992	NN	denotes	domain
T28174	7992-7994	,	denotes	,
T28175	7994-7997	CC	denotes	and
T28176	7998-8006	VBN	denotes	inserted
T28177	8007-8011	IN	denotes	into
T28178	8012-8018	NN	denotes	pcDNA3
T28179	8018-8019	.	denotes	.
T28180	8019-8172	sentence	denotes	We transfected 0.1 μg of reporter plasmid DNA and 2 μg of the expression vector DNA per 6 cm dish into HEK293T cells using Fugene6 transfection reagent.
T28181	8020-8022	PRP	denotes	We
T28182	8023-8034	VBD	denotes	transfected
T28183	8035-8038	CD	denotes	0.1
T28184	8039-8041	NN	denotes	μg
T28185	8042-8044	IN	denotes	of
T28186	8045-8053	NN	denotes	reporter
T28188	8054-8061	NN	denotes	plasmid
T28187	8062-8065	NN	denotes	DNA
T28189	8066-8069	CC	denotes	and
T28190	8070-8071	CD	denotes	2
T28191	8072-8074	NN	denotes	μg
T28192	8075-8077	IN	denotes	of
T28193	8078-8081	DT	denotes	the
T28195	8082-8092	NN	denotes	expression
T28196	8093-8099	NN	denotes	vector
T28194	8100-8103	NN	denotes	DNA
T28197	8104-8107	IN	denotes	per
T28198	8108-8109	CD	denotes	6
T28199	8110-8112	NN	denotes	cm
T28200	8113-8117	NN	denotes	dish
T28201	8118-8122	IN	denotes	into
T28202	8123-8130	NN	denotes	HEK293T
T28203	8131-8136	NNS	denotes	cells
T28204	8137-8142	VBG	denotes	using
T28205	8143-8150	NN	denotes	Fugene6
T28207	8151-8163	NN	denotes	transfection
T28206	8164-8171	NN	denotes	reagent
T28208	8171-8172	.	denotes	.
T28209	8172-8230	sentence	denotes	We analyzed luciferase activity 48 hr after transfection.
T28210	8173-8175	PRP	denotes	We
T28211	8176-8184	VBD	denotes	analyzed
T28212	8185-8195	NN	denotes	luciferase
T28213	8196-8204	NN	denotes	activity
T28214	8205-8207	CD	denotes	48
T28215	8208-8210	NN	denotes	hr
T28216	8211-8216	IN	denotes	after
T28217	8217-8229	NN	denotes	transfection
T28218	8229-8230	.	denotes	.
R6551	T23218	T23217	prep	of,Isolation
R6552	T23219	T23220	compound	mouse,cDNA
R6553	T23220	T23218	pobj	cDNA,of
R6554	T23221	T23222	compound	mr,s
R6555	T23222	T23220	compound	s,cDNA
R6556	T23223	T23222	punct	-,s
R6557	T23225	T23226	nsubj	We,used
R6558	T23227	T23228	det	the,method
R6559	T23228	T23226	dobj	method,used
R6560	T23229	T23228	compound	bioinformatics,method
R6561	T23230	T23231	compound	Digital,Display
R6562	T23231	T23228	appos	Display,method
R6563	T23232	T23231	compound	Differential,Display
R6564	T23233	T23234	punct	(,NCBI
R6565	T23234	T23228	parataxis	NCBI,method
R6566	T23235	T23234	punct	", ",NCBI
R6567	T23236	T23234	npadvmod	UniGene,NCBI
R6568	T23237	T23234	punct	),NCBI
R6569	T23238	T23239	aux	to,screen
R6570	T23239	T23226	advcl	screen,used
R6571	T23240	T23241	amod	novel,genes
R6572	T23241	T23239	dobj	genes,screen
R6573	T23242	T23241	compound	mouse,genes
R6574	T23243	T23241	acl	expressed,genes
R6575	T23244	T23243	advmod	preferentially,expressed
R6576	T23245	T23243	prep	in,expressed
R6577	T23246	T23247	det	the,retina
R6578	T23247	T23245	pobj	retina,in
R6579	T23248	T23226	punct	.,used
R6580	T23250	T23251	nsubj	Some,were
R6581	T23252	T23250	prep	of,Some
R6582	T23253	T23254	det	the,clusters
R6583	T23254	T23252	pobj	clusters,of
R6584	T23255	T23250	prep	in,Some
R6585	T23256	T23257	det	the,database
R6586	T23257	T23255	pobj	database,in
R6587	T23258	T23257	compound	UniGene,database
R6588	T23259	T23251	advmod	mainly,were
R6589	T23260	T23251	prep	from,were
R6590	T23261	T23262	nmod	mouse,cDNAs
R6591	T23262	T23260	pobj	cDNAs,from
R6592	T23263	T23262	amod	retinal,cDNAs
R6593	T23264	T23251	punct	.,were
R6594	T23266	T23267	nummod	One,clone
R6595	T23267	T23268	nsubj	clone,has
R6596	T23269	T23267	prep	in,clone
R6597	T23270	T23271	det	these,clusters
R6598	T23271	T23269	pobj	clusters,in
R6599	T23272	T23273	punct	(,Mm.
R6600	T23273	T23267	parataxis	Mm.,clone
R6601	T23274	T23273	punct	#,Mm.
R6602	T23275	T23273	nummod	246385,Mm.
R6603	T23276	T23273	punct	),Mm.
R6604	T23277	T23278	det	a,homology
R6605	T23278	T23268	dobj	homology,has
R6606	T23279	T23278	amod	weak,homology
R6607	T23280	T23278	prep	with,homology
R6608	T23281	T23282	det	the,genes
R6609	T23282	T23280	pobj	genes,with
R6610	T23283	T23282	amod	polyhomeotic,genes
R6611	T23284	T23282	compound	family,genes
R6612	T23285	T23268	punct	.,has
R6613	T23287	T23288	det	A,fragment
R6614	T23288	T23293	nsubjpass	fragment,amplified
R6615	T23289	T23290	nummod	735,bp
R6616	T23290	T23288	compound	bp,fragment
R6617	T23291	T23290	punct	-,bp
R6618	T23292	T23288	compound	cDNA,fragment
R6619	T23294	T23288	prep	of,fragment
R6620	T23295	T23296	det	this,clone
R6621	T23296	T23294	pobj	clone,of
R6622	T23297	T23288	punct	", ",fragment
R6623	T23298	T23288	acl	encoding,fragment
R6624	T23299	T23300	nmod	amino,140
R6625	T23300	T23298	dobj	140,encoding
R6626	T23301	T23300	nmod	acids,140
R6627	T23302	T23303	punct	–,384
R6628	T23303	T23300	prep	384,140
R6629	T23304	T23293	auxpass	was,amplified
R6630	T23305	T23293	prep	by,amplified
R6631	T23306	T23307	compound	RT,PCR
R6632	T23307	T23305	pobj	PCR,by
R6633	T23308	T23307	punct	-,PCR
R6634	T23309	T23293	prep	from,amplified
R6635	T23310	T23311	nmod	mouse,cDNA
R6636	T23311	T23309	pobj	cDNA,from
R6637	T23312	T23311	nmod	P0,cDNA
R6638	T23313	T23311	amod	retinal,cDNA
R6639	T23314	T23293	punct	.,amplified
R6640	T23316	T23317	det	This,fragment
R6641	T23317	T23318	nsubjpass	fragment,used
R6642	T23319	T23318	auxpass	was,used
R6643	T23320	T23318	prep	as,used
R6644	T23321	T23322	det	the,probe
R6645	T23322	T23320	pobj	probe,as
R6646	T23323	T23318	prep	for,used
R6647	T23324	T23325	compound	library,screening
R6648	T23325	T23323	pobj	screening,for
R6649	T23326	T23325	punct	", ",screening
R6650	T23327	T23328	advmod	in,situ
R6651	T23328	T23329	amod	situ,hybridization
R6652	T23329	T23325	conj	hybridization,screening
R6653	T23330	T23329	cc	and,hybridization
R6654	T23331	T23332	compound	Northern,hybridization
R6655	T23332	T23329	conj	hybridization,hybridization
R6656	T23333	T23318	punct	.,used
R6657	T23335	T23336	det	A,library
R6658	T23336	T23343	nsubjpass	library,screened
R6659	T23337	T23336	nmod	mouse,library
R6660	T23338	T23336	nmod	P0,library
R6661	T23339	T23340	punct	-,P3
R6662	T23340	T23338	prep	P3,P0
R6663	T23341	T23336	amod	retinal,library
R6664	T23342	T23336	compound	cDNA,library
R6665	T23344	T23343	auxpass	was,screened
R6666	T23345	T23343	advcl	using,screened
R6667	T23346	T23347	det	this,fragment
R6668	T23347	T23345	dobj	fragment,using
R6669	T23348	T23347	compound	mouse,fragment
R6670	T23349	T23347	compound	cDNA,fragment
R6671	T23350	T23343	punct	.,screened
R6672	T23352	T23353	amod	Positive,clones
R6673	T23353	T23355	nsubjpass	clones,isolated
R6674	T23354	T23353	compound	bacteriophage,clones
R6675	T23356	T23355	auxpass	were,isolated
R6676	T23357	T23355	cc	and,isolated
R6677	T23358	T23359	det	the,fragment
R6678	T23359	T23366	nsubjpass	fragment,inserted
R6679	T23360	T23361	amod	full,length
R6680	T23361	T23359	compound	length,fragment
R6681	T23362	T23361	punct	-,length
R6682	T23363	T23364	compound	mr,s
R6683	T23364	T23359	compound	s,fragment
R6684	T23365	T23364	punct	-,s
R6685	T23366	T23355	conj	inserted,isolated
R6686	T23367	T23366	auxpass	was,inserted
R6687	T23368	T23366	prep	into,inserted
R6688	T23369	T23368	pobj	pBluescriptII,into
R6689	T23370	T23371	punct	(,Stratagene
R6690	T23371	T23366	parataxis	Stratagene,inserted
R6691	T23372	T23371	punct	),Stratagene
R6692	T23373	T23366	punct	.,inserted
R6693	T23375	T23376	compound	DNA,sequencing
R6694	T23376	T23377	nsubjpass	sequencing,performed
R6695	T23378	T23377	auxpass	was,performed
R6696	T23379	T23377	prep	on,performed
R6697	T23380	T23381	det	both,strands
R6698	T23381	T23379	pobj	strands,on
R6699	T23382	T23377	prep	by,performed
R6700	T23383	T23384	det	the,method
R6701	T23384	T23382	pobj	method,by
R6702	T23385	T23384	compound	cycle,method
R6703	T23386	T23384	compound	sequencing,method
R6704	T23387	T23377	punct	.,performed
R6705	T23389	T23390	det	The,sequence
R6706	T23390	T23392	nsubjpass	sequence,deposited
R6707	T23391	T23390	compound	nucleotide,sequence
R6708	T23393	T23390	prep	for,sequence
R6709	T23394	T23395	compound	mr,s
R6710	T23395	T23397	compound	s,gene
R6711	T23396	T23395	punct	-,s
R6712	T23397	T23393	pobj	gene,for
R6713	T23398	T23392	aux	has,deposited
R6714	T23399	T23392	auxpass	been,deposited
R6715	T23400	T23392	prep	in,deposited
R6716	T23401	T23402	det	the,database
R6717	T23402	T23400	pobj	database,in
R6718	T23403	T23402	compound	GenBank,database
R6719	T23404	T23392	prep	under,deposited
R6720	T23405	T23406	nmod	GenBank,AY458844
R6721	T23406	T23404	pobj	AY458844,under
R6722	T23407	T23406	nmod	Accession,AY458844
R6723	T23408	T23406	nmod	Number,AY458844
R6724	T23409	T23406	punct	#,AY458844
R6725	T23410	T23392	punct	.,deposited
R6726	T23508	T23509	advmod	In,situ
R6727	T23509	T23510	amod	situ,hybridization
R6728	T23512	T23513	det	The,fragment
R6729	T23513	T23518	nsubjpass	fragment,used
R6730	T23514	T23515	nummod	735,bp
R6731	T23515	T23513	compound	bp,fragment
R6732	T23516	T23515	punct	-,bp
R6733	T23517	T23513	compound	cDNA,fragment
R6734	T23519	T23513	prep	of,fragment
R6735	T23520	T23521	compound	mr,s
R6736	T23521	T23519	pobj	s,of
R6737	T23522	T23521	punct	-,s
R6738	T23523	T23513	acl	amplified,fragment
R6739	T23524	T23523	prep	by,amplified
R6740	T23525	T23526	compound	RT,PCR
R6741	T23526	T23524	pobj	PCR,by
R6742	T23527	T23526	punct	-,PCR
R6743	T23528	T23518	auxpass	was,used
R6744	T23529	T23518	prep	as,used
R6745	T23530	T23531	det	a,probe
R6746	T23531	T23529	pobj	probe,as
R6747	T23532	T23518	prep	for,used
R6748	T23533	T23534	advmod	in,situ
R6749	T23534	T23535	amod	situ,hybridization
R6750	T23535	T23532	pobj	hybridization,for
R6751	T23536	T23518	punct	.,used
R6752	T23538	T23539	advmod	In,situ
R6753	T23539	T23540	amod	situ,hybridization
R6754	T23540	T23541	nsubjpass	hybridization,performed
R6755	T23542	T23541	auxpass	was,performed
R6756	T23543	T23544	mark	as,described
R6757	T23544	T23541	advcl	described,performed
R6758	T23545	T23544	advmod	previously,described
R6759	T23546	T23547	punct	[,49
R6760	T23547	T23541	parataxis	49,performed
R6761	T23548	T23547	punct	],49
R6762	T23549	T23541	punct	.,performed
R6763	T23802	T23803	compound	Cell,culture
R6764	T23804	T23803	cc	and,culture
R6765	T23805	T23803	conj	transfection,culture
R6766	T23807	T23808	compound	HEK293T,cells
R6767	T23808	T23809	nsubjpass	cells,maintained
R6768	T23810	T23809	auxpass	were,maintained
R6769	T23811	T23809	prep	at,maintained
R6770	T23812	T23813	nummod	37,°C
R6771	T23813	T23811	pobj	°C,at
R6772	T23814	T23809	prep	in,maintained
R6773	T23815	T23816	poss	Dulbecco,medium
R6774	T23816	T23814	pobj	medium,in
R6775	T23817	T23815	case	's,Dulbecco
R6776	T23818	T23816	amod	modified,medium
R6777	T23819	T23816	poss	Eagle,medium
R6778	T23820	T23819	case	's,Eagle
R6779	T23821	T23816	punct	(,medium
R6780	T23822	T23816	appos	DMEM,medium
R6781	T23823	T23816	punct	),medium
R6782	T23824	T23816	acl	supplemented,medium
R6783	T23825	T23824	prep	with,supplemented
R6784	T23826	T23827	nummod	10,%
R6785	T23827	T23828	nmod	%,serum
R6786	T23828	T23825	pobj	serum,with
R6787	T23829	T23830	amod	fetal,bovine
R6788	T23830	T23828	amod	bovine,serum
R6789	T23831	T23832	punct	(,Sigma
R6790	T23832	T23828	parataxis	Sigma,serum
R6791	T23833	T23832	punct	),Sigma
R6792	T23834	T23828	punct	", ",serum
R6793	T23835	T23836	nummod	100,IU
R6794	T23836	T23837	nmod	IU,penicillin
R6795	T23837	T23828	conj	penicillin,serum
R6796	T23838	T23839	punct	/,ml
R6797	T23839	T23836	prep	ml,IU
R6798	T23840	T23837	cc	and,penicillin
R6799	T23841	T23842	nummod	100,μg
R6800	T23842	T23843	nmod	μg,streptomycin
R6801	T23843	T23837	conj	streptomycin,penicillin
R6802	T23844	T23845	punct	/,ml
R6803	T23845	T23842	prep	ml,μg
R6804	T23846	T23809	punct	.,maintained
R6805	T23848	T23849	amod	Transient,transfection
R6806	T23849	T23850	nsubjpass	transfection,carried
R6807	T23851	T23849	prep	of,transfection
R6808	T23852	T23853	compound	HEK293T,cells
R6809	T23853	T23851	pobj	cells,of
R6810	T23854	T23850	auxpass	was,carried
R6811	T23855	T23850	prt	out,carried
R6812	T23856	T23850	advcl	using,carried
R6813	T23857	T23858	compound	calcium,phosphate
R6814	T23858	T23859	compound	phosphate,method
R6815	T23859	T23856	dobj	method,using
R6816	T23860	T23859	cc	or,method
R6817	T23861	T23862	compound	Fugene6,reagent
R6818	T23862	T23859	conj	reagent,method
R6819	T23863	T23862	compound	transfection,reagent
R6820	T23864	T23865	punct	(,Roche
R6821	T23865	T23862	parataxis	Roche,reagent
R6822	T23866	T23865	punct	),Roche
R6823	T23867	T23850	punct	.,carried
R6824	T23869	T23870	compound	Y79,cells
R6825	T23870	T23872	nsubjpass	cells,maintained
R6826	T23871	T23870	compound	retinoblastoma,cells
R6827	T23873	T23872	auxpass	were,maintained
R6828	T23874	T23872	prep	in,maintained
R6829	T23875	T23876	poss	Iscove,medium
R6830	T23876	T23874	pobj	medium,in
R6831	T23877	T23875	case	's,Iscove
R6832	T23878	T23876	amod	modified,medium
R6833	T23879	T23876	poss	Dulbecco,medium
R6834	T23880	T23879	case	's,Dulbecco
R6835	T23881	T23876	prep	with,medium
R6836	T23882	T23883	nummod	4,mM
R6837	T23883	T23884	compound	mM,glutamine
R6838	T23884	T23881	pobj	glutamine,with
R6839	T23885	T23884	compound	L,glutamine
R6840	T23886	T23884	punct	-,glutamine
R6841	T23887	T23884	acl	adjusted,glutamine
R6842	T23888	T23889	aux	to,contain
R6843	T23889	T23887	advcl	contain,adjusted
R6844	T23890	T23891	nummod	1.5,g
R6845	T23891	T23892	nmod	g,bicarbonate
R6846	T23892	T23889	dobj	bicarbonate,contain
R6847	T23893	T23894	punct	/,L
R6848	T23894	T23891	prep	L,g
R6849	T23895	T23892	compound	sodium,bicarbonate
R6850	T23896	T23892	punct	", ",bicarbonate
R6851	T23897	T23898	nummod	20,%
R6852	T23898	T23899	compound	%,FBS
R6853	T23899	T23892	npadvmod	FBS,bicarbonate
R6854	T23900	T23872	punct	.,maintained
R6855	T23902	T23903	amod	Transient,transfection
R6856	T23903	T23904	nsubjpass	transfection,carried
R6857	T23905	T23904	auxpass	was,carried
R6858	T23906	T23904	prt	out,carried
R6859	T23907	T23904	advcl	using,carried
R6860	T23908	T23909	compound	TransIt,LT1
R6861	T23909	T23907	dobj	LT1,using
R6862	T23910	T23911	punct	(,Mirus
R6863	T23911	T23909	parataxis	Mirus,LT1
R6864	T23912	T23911	punct	),Mirus
R6865	T23913	T23904	punct	.,carried
R6866	T24169	T24170	compound	Northern,blot
R6867	T24170	T24172	compound	blot,analysis
R6868	T24171	T24170	punct	-,blot
R6869	T24174	T24175	nsubjpass	RNA,extracted
R6870	T24176	T24175	auxpass	was,extracted
R6871	T24177	T24175	prep	from,extracted
R6872	T24178	T24179	det	the,tissues
R6873	T24179	T24177	pobj	tissues,from
R6874	T24180	T24179	prep	of,tissues
R6875	T24181	T24182	amod	adult,mice
R6876	T24182	T24180	pobj	mice,of
R6877	T24183	T24175	advcl	using,extracted
R6878	T24184	T24183	dobj	Trizol,using
R6879	T24185	T24186	punct	(,Invitrogen
R6880	T24186	T24184	parataxis	Invitrogen,Trizol
R6881	T24187	T24186	punct	),Invitrogen
R6882	T24188	T24175	punct	.,extracted
R6883	T24190	T24191	det	A,μg
R6884	T24191	T24193	nsubjpass	μg,electrophoresed
R6885	T24192	T24191	nummod	5,μg
R6886	T24194	T24191	prep	of,μg
R6887	T24195	T24196	amod	total,RNA
R6888	T24196	T24194	pobj	RNA,of
R6889	T24197	T24193	auxpass	was,electrophoresed
R6890	T24198	T24193	prep	in,electrophoresed
R6891	T24199	T24200	det	a,gel
R6892	T24200	T24198	pobj	gel,in
R6893	T24201	T24202	nummod	1.0,%
R6894	T24202	T24200	compound	%,gel
R6895	T24203	T24204	compound	agarose,formaldehyde
R6896	T24204	T24200	compound	formaldehyde,gel
R6897	T24205	T24204	punct	-,formaldehyde
R6898	T24206	T24193	cc	and,electrophoresed
R6899	T24207	T24193	conj	transferred,electrophoresed
R6900	T24208	T24207	prep	to,transferred
R6901	T24209	T24210	det	a,membrane
R6902	T24210	T24208	pobj	membrane,to
R6903	T24211	T24210	compound	nylon,membrane
R6904	T24212	T24213	punct	(,GT
R6905	T24213	T24210	parataxis	GT,membrane
R6906	T24214	T24215	compound	Zeta,Probe
R6907	T24215	T24213	compound	Probe,GT
R6908	T24216	T24215	punct	-,Probe
R6909	T24217	T24213	punct	", ",GT
R6910	T24218	T24219	compound	Bio,Rad
R6911	T24219	T24213	npadvmod	Rad,GT
R6912	T24220	T24219	punct	-,Rad
R6913	T24221	T24213	punct	),GT
R6914	T24222	T24193	punct	.,electrophoresed
R6915	T24224	T24225	det	The,fragment
R6916	T24225	T24230	nsubjpass	fragment,used
R6917	T24226	T24227	nummod	735,bp
R6918	T24227	T24225	compound	bp,fragment
R6919	T24228	T24227	punct	-,bp
R6920	T24229	T24225	compound	cDNA,fragment
R6921	T24231	T24225	acl	encoding,fragment
R6922	T24232	T24233	det	the,domain
R6923	T24233	T24231	dobj	domain,encoding
R6924	T24234	T24233	compound	SAM,domain
R6925	T24235	T24233	prep	of,domain
R6926	T24236	T24237	compound	mr,s
R6927	T24237	T24235	pobj	s,of
R6928	T24238	T24237	punct	-,s
R6929	T24239	T24230	auxpass	was,used
R6930	T24240	T24230	prep	as,used
R6931	T24241	T24242	det	a,probe
R6932	T24242	T24240	pobj	probe,as
R6933	T24243	T24230	prep	for,used
R6934	T24244	T24243	pobj	hybridization,for
R6935	T24245	T24230	punct	.,used
R6936	T24247	T24248	nsubjpass	Hybridization,performed
R6937	T24249	T24248	auxpass	was,performed
R6938	T24250	T24248	prep	according,performed
R6939	T24251	T24250	prep	to,according
R6940	T24252	T24253	det	the,manufacturer
R6941	T24253	T24254	poss	manufacturer,protocol
R6942	T24254	T24251	pobj	protocol,to
R6943	T24255	T24253	case	's,manufacturer
R6944	T24256	T24248	punct	.,performed
R6945	T24258	T24259	nsubjpass	Washes,performed
R6946	T24260	T24258	prep	with,Washes
R6947	T24261	T24262	amod	increasing,stringency
R6948	T24262	T24260	pobj	stringency,with
R6949	T24263	T24259	auxpass	were,performed
R6950	T24264	T24259	punct	", ",performed
R6951	T24265	T24266	det	the,last
R6952	T24266	T24267	nsubj	last,being
R6953	T24267	T24259	advcl	being,performed
R6954	T24268	T24267	prep	at,being
R6955	T24269	T24270	nummod	50,°C
R6956	T24270	T24268	pobj	°C,at
R6957	T24271	T24267	prep	in,being
R6958	T24272	T24273	nummod	0.1,citrate
R6959	T24273	T24271	pobj	citrate,in
R6960	T24274	T24272	punct	×,0.1
R6961	T24275	T24273	amod	standard,citrate
R6962	T24276	T24273	compound	saline,citrate
R6963	T24277	T24278	punct	/,%
R6964	T24278	T24273	prep	%,citrate
R6965	T24279	T24278	nummod	0.1,%
R6966	T24280	T24281	compound	sodium,sulfate
R6967	T24281	T24278	appos	sulfate,%
R6968	T24282	T24281	compound	dodecyl,sulfate
R6969	T24283	T24281	punct	(,sulfate
R6970	T24284	T24281	appos	SDS,sulfate
R6971	T24285	T24259	punct	),performed
R6972	T24286	T24259	punct	.,performed
R6973	T24536	T24537	compound	RT,PCR
R6974	T24537	T24539	compound	PCR,analysis
R6975	T24538	T24537	punct	-,PCR
R6976	T24541	T24542	amod	Total,RNA
R6977	T24542	T24543	nsubjpass	RNA,isolated
R6978	T24544	T24543	auxpass	was,isolated
R6979	T24545	T24543	prep	from,isolated
R6980	T24546	T24547	det	each,tissue
R6981	T24547	T24545	pobj	tissue,from
R6982	T24548	T24543	advcl	using,isolated
R6983	T24549	T24548	dobj	Trizol,using
R6984	T24550	T24543	punct	.,isolated
R6985	T24552	T24553	det	A,μg
R6986	T24553	T24555	nsubjpass	μg,transcribed
R6987	T24554	T24553	nummod	1,μg
R6988	T24556	T24553	prep	of,μg
R6989	T24557	T24558	amod	total,RNA
R6990	T24558	T24556	pobj	RNA,of
R6991	T24559	T24555	auxpass	was,transcribed
R6992	T24560	T24555	amod	reverse,transcribed
R6993	T24561	T24555	advcl	using,transcribed
R6994	T24562	T24561	dobj	SuperscriptII,using
R6995	T24563	T24564	punct	(,Invitrogen
R6996	T24564	T24562	parataxis	Invitrogen,SuperscriptII
R6997	T24565	T24564	punct	),Invitrogen
R6998	T24566	T24555	punct	.,transcribed
R6999	T24568	T24569	compound	RT,PCR
R7000	T24569	T24571	compound	PCR,primers
R7001	T24570	T24569	punct	-,PCR
R7002	T24571	T24572	nsubj	primers,were
R7003	T24573	T24571	punct	", ",primers
R7004	T24574	T24575	dep	which,span
R7005	T24575	T24571	relcl	span,primers
R7006	T24576	T24575	dobj	introns,span
R7007	T24577	T24571	punct	", ",primers
R7008	T24578	T24571	prep	for,primers
R7009	T24579	T24578	pobj	detection,for
R7010	T24580	T24579	prep	of,detection
R7011	T24581	T24582	compound	mr,s
R7012	T24582	T24584	compound	s,cDNA
R7013	T24583	T24582	punct	-,s
R7014	T24584	T24580	pobj	cDNA,of
R7015	T24585	T24586	nummod	5,TGTCCAGCCCAGCCAACCCAAGGAGACGACA
R7016	T24586	T24572	attr	TGTCCAGCCCAGCCAACCCAAGGAGACGACA,were
R7017	T24587	T24585	punct	',5
R7018	T24588	T24586	punct	-,TGTCCAGCCCAGCCAACCCAAGGAGACGACA
R7019	T24589	T24586	punct	-,TGTCCAGCCCAGCCAACCCAAGGAGACGACA
R7020	T24590	T24586	nummod	3,TGTCCAGCCCAGCCAACCCAAGGAGACGACA
R7021	T24591	T24586	punct	',TGTCCAGCCCAGCCAACCCAAGGAGACGACA
R7022	T24592	T24586	cc	and,TGTCCAGCCCAGCCAACCCAAGGAGACGACA
R7023	T24593	T24594	nummod	5,TGTGGTCTCCTCATCAGTGAAGA
R7024	T24594	T24586	conj	TGTGGTCTCCTCATCAGTGAAGA,TGTCCAGCCCAGCCAACCCAAGGAGACGACA
R7025	T24595	T24593	punct	',5
R7026	T24596	T24594	punct	-,TGTGGTCTCCTCATCAGTGAAGA
R7027	T24597	T24594	punct	-,TGTGGTCTCCTCATCAGTGAAGA
R7028	T24598	T24594	nummod	3,TGTGGTCTCCTCATCAGTGAAGA
R7029	T24599	T24594	punct	',TGTGGTCTCCTCATCAGTGAAGA
R7030	T24600	T24572	punct	.,were
R7031	T24602	T24603	compound	Product,size
R7032	T24603	T24604	nsubj	size,was
R7033	T24605	T24606	nummod	292,bp
R7034	T24606	T24604	attr	bp,was
R7035	T24607	T24608	punct	(,965
R7036	T24608	T24604	parataxis	965,was
R7037	T24609	T24608	nmod	positions,965
R7038	T24610	T24611	punct	–,1256
R7039	T24611	T24608	prep	1256,965
R7040	T24612	T24608	prep	of,965
R7041	T24613	T24614	nmod	Genbank,AY458844
R7042	T24614	T24612	pobj	AY458844,of
R7043	T24615	T24614	nmod	Accession,AY458844
R7044	T24616	T24614	nmod	Number,AY458844
R7045	T24617	T24614	punct	#,AY458844
R7046	T24618	T24608	punct	),965
R7047	T24619	T24604	punct	.,was
R7048	T24621	T24622	compound	Primer,pairs
R7049	T24622	T24623	nsubj	pairs,were
R7050	T24624	T24622	prep	for,pairs
R7051	T24625	T24626	compound	mouse,G3PDH
R7052	T24626	T24624	pobj	G3PDH,for
R7053	T24627	T24628	nummod	5,ACCACAGTCCATGCCATCAC
R7054	T24628	T24623	attr	ACCACAGTCCATGCCATCAC,were
R7055	T24629	T24627	punct	',5
R7056	T24630	T24628	punct	-,ACCACAGTCCATGCCATCAC
R7057	T24631	T24628	punct	-,ACCACAGTCCATGCCATCAC
R7058	T24632	T24628	nummod	3,ACCACAGTCCATGCCATCAC
R7059	T24633	T24628	punct	',ACCACAGTCCATGCCATCAC
R7060	T24634	T24628	cc	and,ACCACAGTCCATGCCATCAC
R7061	T24635	T24636	nummod	5,TCCACCACCCTGTTGCTGTA
R7062	T24636	T24628	conj	TCCACCACCCTGTTGCTGTA,ACCACAGTCCATGCCATCAC
R7063	T24637	T24635	punct	',5
R7064	T24638	T24636	punct	-,TCCACCACCCTGTTGCTGTA
R7065	T24639	T24636	punct	-,TCCACCACCCTGTTGCTGTA
R7066	T24640	T24636	nummod	3,TCCACCACCCTGTTGCTGTA
R7067	T24641	T24636	punct	',TCCACCACCCTGTTGCTGTA
R7068	T24642	T24643	dep	which,amplified
R7069	T24643	T24628	relcl	amplified,ACCACAGTCCATGCCATCAC
R7070	T24644	T24645	det	a,product
R7071	T24645	T24643	dobj	product,amplified
R7072	T24646	T24647	nummod	452,bp
R7073	T24647	T24645	compound	bp,product
R7074	T24648	T24647	punct	-,bp
R7075	T24649	T24650	punct	(,587
R7076	T24650	T24643	parataxis	587,amplified
R7077	T24651	T24650	nmod	positions,587
R7078	T24652	T24653	punct	–,1038
R7079	T24653	T24650	prep	1038,587
R7080	T24654	T24650	prep	of,587
R7081	T24655	T24656	nmod	Genbank,BC85275
R7082	T24656	T24654	pobj	BC85275,of
R7083	T24657	T24656	nmod	Accession,BC85275
R7084	T24658	T24656	nmod	Number,BC85275
R7085	T24659	T24656	punct	#,BC85275
R7086	T24660	T24650	punct	),587
R7087	T24661	T24623	punct	.,were
R7088	T25116	T25117	nmod	Yeast,screening
R7089	T25118	T25119	nummod	two,hybrid
R7090	T25119	T25117	compound	hybrid,screening
R7091	T25120	T25119	punct	-,hybrid
R7092	T25121	T25117	cc	and,screening
R7093	T25122	T25123	compound	GAL4,assay
R7094	T25123	T25117	conj	assay,screening
R7095	T25125	T25126	nsubj	We,carried
R7096	T25127	T25126	prt	out,carried
R7097	T25128	T25129	nmod	yeast,experiments
R7098	T25129	T25126	dobj	experiments,carried
R7099	T25130	T25131	nummod	two,hybrid
R7100	T25131	T25129	compound	hybrid,experiments
R7101	T25132	T25131	punct	-,hybrid
R7102	T25133	T25126	advcl	using,carried
R7103	T25134	T25135	det	the,system
R7104	T25135	T25133	dobj	system,using
R7105	T25136	T25135	nmod	MATCHMAKER,system
R7106	T25137	T25135	nmod	GAL4,system
R7107	T25138	T25139	nummod	two,hybrid
R7108	T25139	T25135	compound	hybrid,system
R7109	T25140	T25139	punct	-,hybrid
R7110	T25141	T25135	nummod	3,system
R7111	T25142	T25143	punct	(,Bioscience
R7112	T25143	T25135	parataxis	Bioscience,system
R7113	T25144	T25143	compound	BD,Bioscience
R7114	T25145	T25143	punct	),Bioscience
R7115	T25146	T25147	mark	as,recommended
R7116	T25147	T25126	advcl	recommended,carried
R7117	T25148	T25147	agent	by,recommended
R7118	T25149	T25150	det	the,manufacturer
R7119	T25150	T25148	pobj	manufacturer,by
R7120	T25151	T25126	punct	.,carried
R7121	T25153	T25154	nsubj	We,cloned
R7122	T25155	T25156	det	the,s
R7123	T25156	T25154	dobj	s,cloned
R7124	T25157	T25158	amod	full,length
R7125	T25158	T25156	compound	length,s
R7126	T25159	T25158	punct	-,length
R7127	T25160	T25156	compound	mr,s
R7128	T25161	T25156	punct	-,s
R7129	T25162	T25154	prep	into,cloned
R7130	T25163	T25164	det	the,vector
R7131	T25164	T25162	pobj	vector,into
R7132	T25165	T25164	compound	pGBKT7,vector
R7133	T25166	T25154	cc	and,cloned
R7134	T25167	T25154	conj	used,cloned
R7135	T25168	T25167	dobj	it,used
R7136	T25169	T25170	aux	to,screen
R7137	T25170	T25167	advcl	screen,used
R7138	T25171	T25172	det	a,library
R7139	T25172	T25170	dobj	library,screen
R7140	T25173	T25172	prep	of,library
R7141	T25174	T25175	nmod	mouse,cDNAs
R7142	T25175	T25173	pobj	cDNAs,of
R7143	T25176	T25175	nmod	P0,cDNAs
R7144	T25177	T25178	punct	-,P3
R7145	T25178	T25176	prep	P3,P0
R7146	T25179	T25175	amod	retinal,cDNAs
R7147	T25180	T25175	prep	in,cDNAs
R7148	T25181	T25182	det	the,vector
R7149	T25182	T25180	pobj	vector,in
R7150	T25183	T25182	compound	pGADT7,vector
R7151	T25184	T25154	punct	.,cloned
R7152	T25186	T25187	nsubj	Transformants,picked
R7153	T25188	T25189	dep	that,conferred
R7154	T25189	T25186	relcl	conferred,Transformants
R7155	T25190	T25189	dobj	growth,conferred
R7156	T25191	T25187	aux	were,picked
R7157	T25192	T25187	punct	", ",picked
R7158	T25193	T25187	conj	isolated,picked
R7159	T25194	T25193	cc	and,isolated
R7160	T25195	T25193	conj	re-introduced,isolated
R7161	T25196	T25195	prep	with,re-introduced
R7162	T25197	T25198	det	the,bait
R7163	T25198	T25196	pobj	bait,with
R7164	T25199	T25195	prep	into,re-introduced
R7165	T25200	T25199	pobj	AH109,into
R7166	T25201	T25202	aux	to,confirm
R7167	T25202	T25187	advcl	confirm,picked
R7168	T25203	T25202	dobj	interaction,confirm
R7169	T25204	T25187	punct	.,picked
R7170	T25206	T25207	compound	Plasmid,DNA
R7171	T25207	T25208	nsubjpass	DNA,isolated
R7172	T25209	T25208	auxpass	was,isolated
R7173	T25210	T25208	prep	from,isolated
R7174	T25211	T25210	pobj	yeast,from
R7175	T25212	T25208	advcl	using,isolated
R7176	T25213	T25214	compound	RPM,Kit
R7177	T25214	T25212	dobj	Kit,using
R7178	T25215	T25216	compound	Yeast,Plasmid
R7179	T25216	T25217	compound	Plasmid,Isolation
R7180	T25217	T25214	compound	Isolation,Kit
R7181	T25218	T25219	punct	(,Qbiogene
R7182	T25219	T25214	parataxis	Qbiogene,Kit
R7183	T25220	T25219	punct	),Qbiogene
R7184	T25221	T25208	punct	.,isolated
R7185	T25223	T25224	prep	In,picked
R7186	T25225	T25223	pobj	order,In
R7187	T25226	T25227	aux	to,confirm
R7188	T25227	T25225	acl	confirm,order
R7189	T25228	T25229	det	the,interactions
R7190	T25229	T25227	dobj	interactions,confirm
R7191	T25230	T25229	compound	protein,interactions
R7192	T25231	T25224	punct	", ",picked
R7193	T25232	T25233	amod	single,colonies
R7194	T25233	T25224	nsubjpass	colonies,picked
R7195	T25234	T25224	auxpass	were,picked
R7196	T25235	T25224	cc	and,picked
R7197	T25236	T25224	conj	grown,picked
R7198	T25237	T25236	advmod	individually,grown
R7199	T25238	T25236	prep	in,grown
R7200	T25239	T25240	amod	synthetic,media
R7201	T25240	T25238	pobj	media,in
R7202	T25241	T25240	amod	complete,media
R7203	T25242	T25240	acl	lacking,media
R7204	T25243	T25244	nmod	leucine,tryptophan
R7205	T25244	T25242	dobj	tryptophan,lacking
R7206	T25245	T25244	punct	", ",tryptophan
R7207	T25246	T25242	cc	and,lacking
R7208	T25247	T25242	conj	containing,lacking
R7209	T25248	T25249	compound	X,gal
R7210	T25249	T25247	dobj	gal,containing
R7211	T25250	T25249	punct	-,gal
R7212	T25251	T25224	punct	.,picked
R7213	T25253	T25254	prep	After,selected
R7214	T25255	T25256	quantmod	4,5
R7215	T25256	T25258	nummod	5,days
R7216	T25257	T25256	punct	–,5
R7217	T25258	T25253	pobj	days,After
R7218	T25259	T25254	punct	", ",selected
R7219	T25260	T25261	compound	X,gal
R7220	T25261	T25263	npadvmod	gal,positive
R7221	T25262	T25261	compound	-,gal
R7222	T25263	T25264	amod	positive,clones
R7223	T25264	T25254	nsubjpass	clones,selected
R7224	T25265	T25254	auxpass	were,selected
R7225	T25266	T25254	cc	and,selected
R7226	T25267	T25254	conj	analyzed,selected
R7227	T25268	T25254	punct	.,selected
R7228	T25270	T25271	aux	To,test
R7229	T25271	T25272	advcl	test,subcloned
R7230	T25273	T25274	compound	self,interaction
R7231	T25274	T25271	dobj	interaction,test
R7232	T25275	T25274	punct	-,interaction
R7233	T25276	T25274	prep	of,interaction
R7234	T25277	T25278	compound	mr,s
R7235	T25278	T25276	pobj	s,of
R7236	T25279	T25278	punct	-,s
R7237	T25280	T25272	punct	", ",subcloned
R7238	T25281	T25272	nsubj	we,subcloned
R7239	T25282	T25283	det	the,length
R7240	T25283	T25272	dobj	length,subcloned
R7241	T25284	T25283	amod	full,length
R7242	T25285	T25283	punct	-,length
R7243	T25286	T25283	punct	", ",length
R7244	T25287	T25288	compound	N,terminus
R7245	T25288	T25283	conj	terminus,length
R7246	T25289	T25288	punct	-,terminus
R7247	T25290	T25291	punct	(,1
R7248	T25291	T25288	parataxis	1,terminus
R7249	T25292	T25291	nmod	amino,1
R7250	T25293	T25291	nmod	acids,1
R7251	T25294	T25295	punct	–,400
R7252	T25295	T25291	prep	400,1
R7253	T25296	T25291	punct	),1
R7254	T25297	T25288	cc	and,terminus
R7255	T25298	T25299	compound	C,terminus
R7256	T25299	T25288	conj	terminus,terminus
R7257	T25300	T25299	punct	-,terminus
R7258	T25301	T25302	punct	(,391
R7259	T25302	T25299	parataxis	391,terminus
R7260	T25303	T25302	nmod	amino,391
R7261	T25304	T25302	nmod	acids,391
R7262	T25305	T25306	punct	–,542
R7263	T25306	T25302	prep	542,391
R7264	T25307	T25302	punct	),391
R7265	T25308	T25283	prep	of,length
R7266	T25309	T25310	compound	mr,s
R7267	T25310	T25308	pobj	s,of
R7268	T25311	T25310	punct	-,s
R7269	T25312	T25272	prep	into,subcloned
R7270	T25313	T25314	nmod	pGBKT7,vectors
R7271	T25314	T25312	pobj	vectors,into
R7272	T25315	T25313	cc	and,pGBKT7
R7273	T25316	T25313	conj	pGADT7,pGBKT7
R7274	T25317	T25272	punct	.,subcloned
R7275	T25319	T25320	nsubj	We,assessed
R7276	T25321	T25320	dobj	interactions,assessed
R7277	T25322	T25320	prep	by,assessed
R7278	T25323	T25322	pcomp	scoring,by
R7279	T25324	T25325	amod	blue,color
R7280	T25325	T25323	dobj	color,scoring
R7281	T25326	T25323	prep	on,scoring
R7282	T25327	T25326	pobj	plates,on
R7283	T25328	T25327	prep	of,plates
R7284	T25329	T25328	pobj	medium,of
R7285	T25330	T25327	acl	containing,plates
R7286	T25331	T25332	compound	X,gal
R7287	T25332	T25330	dobj	gal,containing
R7288	T25333	T25332	punct	-,gal
R7289	T25334	T25320	punct	.,assessed
R7290	T26198	T26199	compound	Immunoprecipitation,assay
R7291	T26201	T26202	prep	For,subcloned
R7292	T26203	T26204	compound	immunoprecipitation,assay
R7293	T26204	T26201	pobj	assay,For
R7294	T26205	T26202	punct	", ",subcloned
R7295	T26206	T26207	nummod	4,hemagglutinin
R7296	T26207	T26209	npadvmod	hemagglutinin,tagged
R7297	T26208	T26206	punct	×,4
R7298	T26209	T26217	amod	tagged,fragment
R7299	T26210	T26207	punct	(,hemagglutinin
R7300	T26211	T26207	appos	HA,hemagglutinin
R7301	T26212	T26207	punct	),hemagglutinin
R7302	T26213	T26207	cc	or,hemagglutinin
R7303	T26214	T26215	nummod	3,Flag
R7304	T26215	T26207	conj	Flag,hemagglutinin
R7305	T26216	T26214	punct	×,3
R7306	T26217	T26202	nsubjpass	fragment,subcloned
R7307	T26218	T26217	compound	cDNA,fragment
R7308	T26219	T26217	acl	encoding,fragment
R7309	T26220	T26221	amod	full,length
R7310	T26221	T26223	compound	length,s
R7311	T26222	T26221	punct	-,length
R7312	T26223	T26219	dobj	s,encoding
R7313	T26224	T26223	compound	mr,s
R7314	T26225	T26223	punct	-,s
R7315	T26226	T26227	punct	(,HA
R7316	T26227	T26223	parataxis	HA,s
R7317	T26228	T26227	amod	full,HA
R7318	T26229	T26227	punct	-,HA
R7319	T26230	T26227	cc	and,HA
R7320	T26231	T26232	compound	Flag,mrs
R7321	T26232	T26227	conj	mrs,HA
R7322	T26233	T26232	punct	-,mrs
R7323	T26234	T26227	punct	),HA
R7324	T26235	T26223	punct	", ",s
R7325	T26236	T26237	det	a,fragment
R7326	T26237	T26223	conj	fragment,s
R7327	T26238	T26237	compound	cDNA,fragment
R7328	T26239	T26237	acl	encoding,fragment
R7329	T26240	T26241	nmod	amino,1
R7330	T26241	T26239	dobj	1,encoding
R7331	T26242	T26241	nmod	acids,1
R7332	T26243	T26244	punct	–,400
R7333	T26244	T26241	prep	400,1
R7334	T26245	T26246	punct	(,HA
R7335	T26246	T26237	parataxis	HA,fragment
R7336	T26247	T26246	compound	ΔSAM,HA
R7337	T26248	T26246	punct	-,HA
R7338	T26249	T26246	cc	and,HA
R7339	T26250	T26251	compound	Flag,ΔSAM
R7340	T26251	T26246	conj	ΔSAM,HA
R7341	T26252	T26251	punct	-,ΔSAM
R7342	T26253	T26246	punct	),HA
R7343	T26254	T26237	punct	", ",fragment
R7344	T26255	T26237	cc	and,fragment
R7345	T26256	T26257	det	a,fragment
R7346	T26257	T26237	conj	fragment,fragment
R7347	T26258	T26257	compound	cDNA,fragment
R7348	T26259	T26257	acl	encoding,fragment
R7349	T26260	T26261	nmod	amino,400
R7350	T26261	T26259	dobj	400,encoding
R7351	T26262	T26261	nmod	acids,400
R7352	T26263	T26264	punct	–,542
R7353	T26264	T26261	prep	542,400
R7354	T26265	T26266	punct	(,SAM
R7355	T26266	T26257	parataxis	SAM,fragment
R7356	T26267	T26266	compound	Flag,SAM
R7357	T26268	T26266	punct	-,SAM
R7358	T26269	T26266	punct	),SAM
R7359	T26270	T26202	auxpass	were,subcloned
R7360	T26271	T26202	prep	into,subcloned
R7361	T26272	T26273	det	the,vector
R7362	T26273	T26271	pobj	vector,into
R7363	T26274	T26273	nmod	pcDNA3,vector
R7364	T26275	T26276	punct	(,Invitrogen
R7365	T26276	T26274	parataxis	Invitrogen,pcDNA3
R7366	T26277	T26276	punct	),Invitrogen
R7367	T26278	T26273	compound	expression,vector
R7368	T26279	T26202	punct	.,subcloned
R7369	T26281	T26282	nsubj	We,constructed
R7370	T26283	T26282	advmod	also,constructed
R7371	T26284	T26285	nummod	two,mutants
R7372	T26285	T26282	dobj	mutants,constructed
R7373	T26286	T26287	npadvmod	site,directed
R7374	T26287	T26285	amod	directed,mutants
R7375	T26288	T26287	punct	-,directed
R7376	T26289	T26285	punct	", ",mutants
R7377	T26290	T26291	compound	Flag,W404A
R7378	T26291	T26285	appos	W404A,mutants
R7379	T26292	T26291	punct	-,W404A
R7380	T26293	T26291	cc	and,W404A
R7381	T26294	T26295	compound	Flag,G453A
R7382	T26295	T26291	conj	G453A,W404A
R7383	T26296	T26295	punct	-,G453A
R7384	T26297	T26282	punct	", ",constructed
R7385	T26298	T26282	advcl	using,constructed
R7386	T26299	T26298	dobj	PCR,using
R7387	T26300	T26282	punct	.,constructed
R7388	T26302	T26303	nsubjpass	Each,introduced
R7389	T26304	T26302	prep	of,Each
R7390	T26305	T26306	det	these,mutations
R7391	T26306	T26304	pobj	mutations,of
R7392	T26307	T26303	auxpass	was,introduced
R7393	T26308	T26303	advmod	also,introduced
R7394	T26309	T26303	prep	into,introduced
R7395	T26310	T26309	pobj	Flag,into
R7396	T26311	T26310	punct	-,Flag
R7397	T26312	T26313	compound	mr,s
R7398	T26313	T26310	appos	s,Flag
R7399	T26314	T26313	punct	-,s
R7400	T26315	T26303	punct	.,introduced
R7401	T26317	T26318	nsubj	We,transfected
R7402	T26319	T26320	compound	HEK293T,cells
R7403	T26320	T26318	dobj	cells,transfected
R7404	T26321	T26318	prep	with,transfected
R7405	T26322	T26323	nummod	5,μg
R7406	T26323	T26321	pobj	μg,with
R7407	T26324	T26323	prep	of,μg
R7408	T26325	T26326	compound	plasmid,DNA
R7409	T26326	T26324	pobj	DNA,of
R7410	T26327	T26323	prep	per,μg
R7411	T26328	T26329	nummod	6,cm
R7412	T26329	T26330	compound	cm,dish
R7413	T26330	T26327	pobj	dish,per
R7414	T26331	T26318	prep	by,transfected
R7415	T26332	T26333	compound	calcium,phosphate
R7416	T26333	T26334	compound	phosphate,method
R7417	T26334	T26331	pobj	method,by
R7418	T26335	T26318	punct	.,transfected
R7419	T26337	T26338	advmod	Approximately,48
R7420	T26338	T26339	nummod	48,hr
R7421	T26339	T26340	npadvmod	hr,after
R7422	T26340	T26341	prep	after,harvested
R7423	T26342	T26340	pobj	transfection,after
R7424	T26343	T26341	punct	", ",harvested
R7425	T26344	T26341	nsubjpass	cells,harvested
R7426	T26345	T26341	auxpass	were,harvested
R7427	T26346	T26341	prep	in,harvested
R7428	T26347	T26348	compound	immunoprecipitation,buffer
R7429	T26348	T26346	pobj	buffer,in
R7430	T26349	T26350	punct	(,HCl
R7431	T26350	T26341	parataxis	HCl,harvested
R7432	T26351	T26352	nummod	50,mM
R7433	T26352	T26350	compound	mM,HCl
R7434	T26353	T26350	compound	Tris,HCl
R7435	T26354	T26350	punct	-,HCl
R7436	T26355	T26356	punct	[,pH
R7437	T26356	T26350	parataxis	pH,HCl
R7438	T26357	T26356	nummod	7.5,pH
R7439	T26358	T26356	punct	],pH
R7440	T26359	T26350	punct	", ",HCl
R7441	T26360	T26361	nummod	1,mM
R7442	T26361	T26362	compound	mM,EDTA
R7443	T26362	T26350	appos	EDTA,HCl
R7444	T26363	T26350	punct	", ",HCl
R7445	T26364	T26365	nummod	2,mM
R7446	T26365	T26366	compound	mM,MgCl2
R7447	T26366	T26350	appos	MgCl2,HCl
R7448	T26367	T26350	punct	", ",HCl
R7449	T26368	T26369	nummod	150,mM
R7450	T26369	T26370	compound	mM,NaCl
R7451	T26370	T26350	appos	NaCl,HCl
R7452	T26371	T26350	punct	", ",HCl
R7453	T26372	T26373	nummod	1,mM
R7454	T26373	T26374	compound	mM,fluoride
R7455	T26374	T26350	appos	fluoride,HCl
R7456	T26375	T26374	compound	phenylmethylsulfonyl,fluoride
R7457	T26376	T26377	punct	[,Wako
R7458	T26377	T26374	parataxis	Wako,fluoride
R7459	T26378	T26377	punct	],Wako
R7460	T26379	T26350	punct	", ",HCl
R7461	T26380	T26381	nummod	1,%
R7462	T26381	T26382	compound	%,P
R7463	T26382	T26350	appos	P,HCl
R7464	T26383	T26382	compound	Nonidet,P
R7465	T26384	T26382	punct	-,P
R7466	T26385	T26382	nummod	40,P
R7467	T26386	T26350	punct	", ",HCl
R7468	T26387	T26388	nummod	10,%
R7469	T26388	T26389	compound	%,glycerol
R7470	T26389	T26350	appos	glycerol,HCl
R7471	T26390	T26350	punct	),HCl
R7472	T26391	T26341	prep	in,harvested
R7473	T26392	T26393	det	the,presence
R7474	T26393	T26391	pobj	presence,in
R7475	T26394	T26393	prep	of,presence
R7476	T26395	T26396	compound	protease,inhibitor
R7477	T26396	T26397	compound	inhibitor,tablets
R7478	T26397	T26394	pobj	tablets,of
R7479	T26398	T26397	compound	cocktail,tablets
R7480	T26399	T26400	punct	(,Roche
R7481	T26400	T26397	parataxis	Roche,tablets
R7482	T26401	T26400	punct	),Roche
R7483	T26402	T26341	punct	.,harvested
R7484	T26404	T26405	prep	For,mixed
R7485	T26406	T26407	det	each,reaction
R7486	T26407	T26404	pobj	reaction,For
R7487	T26408	T26405	punct	", ",mixed
R7488	T26409	T26410	nummod	1,mg
R7489	T26410	T26405	nsubjpass	mg,mixed
R7490	T26411	T26410	prep	of,mg
R7491	T26412	T26413	compound	cell,lysate
R7492	T26413	T26411	pobj	lysate,of
R7493	T26414	T26405	auxpass	was,mixed
R7494	T26415	T26405	prep	with,mixed
R7495	T26416	T26417	nummod	0.5,μg
R7496	T26417	T26415	pobj	μg,with
R7497	T26418	T26417	prep	of,μg
R7498	T26419	T26420	amod	anti-Flag,antibody
R7499	T26420	T26418	pobj	antibody,of
R7500	T26421	T26422	punct	(,F3165
R7501	T26422	T26420	parataxis	F3165,antibody
R7502	T26423	T26422	nmod	SIGMA,F3165
R7503	T26424	T26422	punct	", ",F3165
R7504	T26425	T26422	punct	),F3165
R7505	T26426	T26417	cc	and,μg
R7506	T26427	T26428	nummod	15,μl
R7507	T26428	T26417	conj	μl,μg
R7508	T26429	T26428	prep	of,μl
R7509	T26430	T26431	compound	protein,Sepharose
R7510	T26431	T26429	pobj	Sepharose,of
R7511	T26432	T26431	compound	G,Sepharose
R7512	T26433	T26431	punct	-,Sepharose
R7513	T26434	T26435	punct	(,Amersham
R7514	T26435	T26431	parataxis	Amersham,Sepharose
R7515	T26436	T26435	punct	),Amersham
R7516	T26437	T26405	prep	on,mixed
R7517	T26438	T26439	det	a,wheel
R7518	T26439	T26437	pobj	wheel,on
R7519	T26440	T26439	amod	rotating,wheel
R7520	T26441	T26405	prep	at,mixed
R7521	T26442	T26443	nummod	4,°C
R7522	T26443	T26441	pobj	°C,at
R7523	T26444	T26405	prep	for,mixed
R7524	T26445	T26446	nummod	2,hrs
R7525	T26446	T26444	pobj	hrs,for
R7526	T26447	T26405	punct	.,mixed
R7527	T26449	T26450	compound	Protein,concentration
R7528	T26450	T26451	nsubjpass	concentration,determined
R7529	T26452	T26451	auxpass	was,determined
R7530	T26453	T26451	prep	by,determined
R7531	T26454	T26455	det	the,system
R7532	T26455	T26453	pobj	system,by
R7533	T26456	T26457	compound	BCA,assay
R7534	T26457	T26455	compound	assay,system
R7535	T26458	T26457	compound	protein,assay
R7536	T26459	T26460	punct	(,Pierce
R7537	T26460	T26455	parataxis	Pierce,system
R7538	T26461	T26460	punct	),Pierce
R7539	T26462	T26451	punct	.,determined
R7540	T26464	T26465	det	The,beads
R7541	T26465	T26466	nsubjpass	beads,washed
R7542	T26467	T26466	auxpass	were,washed
R7543	T26468	T26466	advmod	then,washed
R7544	T26469	T26470	nummod	three,times
R7545	T26470	T26466	npadvmod	times,washed
R7546	T26471	T26466	prep	with,washed
R7547	T26472	T26473	compound	immunoprecipitation,buffer
R7548	T26473	T26471	pobj	buffer,with
R7549	T26474	T26466	cc	and,washed
R7550	T26475	T26466	conj	followed,washed
R7551	T26476	T26475	agent	by,followed
R7552	T26477	T26478	nummod	three,times
R7553	T26478	T26476	pobj	times,by
R7554	T26479	T26478	prep	with,times
R7555	T26480	T26481	compound	wash,buffer
R7556	T26481	T26479	pobj	buffer,with
R7557	T26482	T26483	punct	(,HCl
R7558	T26483	T26481	parataxis	HCl,buffer
R7559	T26484	T26485	nummod	50,mM
R7560	T26485	T26483	compound	mM,HCl
R7561	T26486	T26483	compound	Tris,HCl
R7562	T26487	T26483	punct	-,HCl
R7563	T26488	T26489	punct	[,pH
R7564	T26489	T26483	parataxis	pH,HCl
R7565	T26490	T26489	nummod	7.5,pH
R7566	T26491	T26489	punct	],pH
R7567	T26492	T26483	punct	", ",HCl
R7568	T26493	T26494	nummod	150,mM
R7569	T26494	T26495	compound	mM,NaCl
R7570	T26495	T26483	appos	NaCl,HCl
R7571	T26496	T26483	punct	", ",HCl
R7572	T26497	T26498	nummod	20,mM
R7573	T26498	T26499	compound	mM,MgCl2
R7574	T26499	T26483	appos	MgCl2,HCl
R7575	T26500	T26483	punct	),HCl
R7576	T26501	T26466	punct	.,washed
R7577	T26503	T26504	nsubjpass	Proteins,boiled
R7578	T26505	T26504	auxpass	were,boiled
R7579	T26506	T26504	advmod	then,boiled
R7580	T26507	T26504	prep	for,boiled
R7581	T26508	T26509	nummod	5,min
R7582	T26509	T26507	pobj	min,for
R7583	T26510	T26504	prep	in,boiled
R7584	T26511	T26512	compound	SDS,buffer
R7585	T26512	T26510	pobj	buffer,in
R7586	T26513	T26512	compound	sample,buffer
R7587	T26514	T26515	punct	(,SDS
R7588	T26515	T26512	parataxis	SDS,buffer
R7589	T26516	T26517	nummod	1,%
R7590	T26517	T26515	compound	%,SDS
R7591	T26518	T26515	punct	", ",SDS
R7592	T26519	T26520	nummod	1,mM
R7593	T26520	T26521	compound	mM,Tris
R7594	T26521	T26515	appos	Tris,SDS
R7595	T26522	T26523	punct	[,pH
R7596	T26523	T26521	parataxis	pH,Tris
R7597	T26524	T26523	nummod	6.8,pH
R7598	T26525	T26523	punct	],pH
R7599	T26526	T26515	punct	", ",SDS
R7600	T26527	T26528	nummod	40,mM
R7601	T26528	T26529	compound	mM,DTT
R7602	T26529	T26515	appos	DTT,SDS
R7603	T26530	T26515	punct	", ",SDS
R7604	T26531	T26532	nummod	4,%
R7605	T26532	T26533	compound	%,glycerol
R7606	T26533	T26515	appos	glycerol,SDS
R7607	T26534	T26515	punct	", ",SDS
R7608	T26535	T26536	nummod	0.01,%
R7609	T26536	T26537	compound	%,Y
R7610	T26537	T26515	appos	Y,SDS
R7611	T26538	T26537	compound	pyronine,Y
R7612	T26539	T26515	punct	),SDS
R7613	T26540	T26504	punct	.,boiled
R7614	T26542	T26543	det	The,supernatants
R7615	T26543	T26544	nsubjpass	supernatants,fractionated
R7616	T26545	T26544	auxpass	were,fractionated
R7617	T26546	T26544	prep	by,fractionated
R7618	T26547	T26548	nmod	sodium,sulfate
R7619	T26548	T26550	nmod	sulfate,gel
R7620	T26549	T26548	nmod	dodecyl,sulfate
R7621	T26550	T26553	compound	gel,electrophoresis
R7622	T26551	T26548	punct	-,sulfate
R7623	T26552	T26548	appos	polyacrylamide,sulfate
R7624	T26553	T26546	pobj	electrophoresis,by
R7625	T26554	T26555	punct	(,PAGE
R7626	T26555	T26553	parataxis	PAGE,electrophoresis
R7627	T26556	T26555	compound	SDS,PAGE
R7628	T26557	T26555	punct	-,PAGE
R7629	T26558	T26555	punct	),PAGE
R7630	T26559	T26544	cc	and,fractionated
R7631	T26560	T26544	conj	transferred,fractionated
R7632	T26561	T26560	prep	to,transferred
R7633	T26562	T26563	det	the,membrane
R7634	T26563	T26561	pobj	membrane,to
R7635	T26564	T26563	compound	nitrocellulose,membrane
R7636	T26565	T26566	punct	(,Medium
R7637	T26566	T26563	parataxis	Medium,membrane
R7638	T26567	T26568	compound	Trans,Blot
R7639	T26568	T26566	compound	Blot,Medium
R7640	T26569	T26568	punct	-,Blot
R7641	T26570	T26566	compound	Transfer,Medium
R7642	T26571	T26566	punct	", ",Medium
R7643	T26572	T26573	compound	Bio,Rad
R7644	T26573	T26566	appos	Rad,Medium
R7645	T26574	T26573	punct	-,Rad
R7646	T26575	T26566	punct	),Medium
R7647	T26576	T26544	punct	.,fractionated
R7648	T26578	T26579	compound	Western,blotting
R7649	T26579	T26580	nsubjpass	blotting,performed
R7650	T26581	T26580	auxpass	was,performed
R7651	T26582	T26580	prep	with,performed
R7652	T26583	T26584	nmod	rabbit,antibody
R7653	T26584	T26582	pobj	antibody,with
R7654	T26585	T26584	amod	polyclonal,antibody
R7655	T26586	T26584	amod	anti-HA,antibody
R7656	T26587	T26588	punct	(,Cruz
R7657	T26588	T26584	parataxis	Cruz,antibody
R7658	T26589	T26588	compound	Santa,Cruz
R7659	T26590	T26588	punct	", ",Cruz
R7660	T26591	T26592	punct	#,sc
R7661	T26592	T26588	appos	sc,Cruz
R7662	T26593	T26592	punct	-,sc
R7663	T26594	T26592	nummod	805,sc
R7664	T26595	T26588	punct	),Cruz
R7665	T26596	T26580	punct	.,performed
R7666	T26598	T26599	nsubjpass	Signals,detected
R7667	T26600	T26599	auxpass	were,detected
R7668	T26601	T26599	prep	with,detected
R7669	T26602	T26603	compound	horseradish,peroxidase
R7670	T26603	T26604	npadvmod	peroxidase,conjugated
R7671	T26604	T26606	amod	conjugated,IgG
R7672	T26605	T26604	punct	-,conjugated
R7673	T26606	T26601	pobj	IgG,with
R7674	T26607	T26606	nmod	goat,IgG
R7675	T26608	T26606	amod	anti-rabbit,IgG
R7676	T26609	T26606	cc	and,IgG
R7677	T26610	T26611	nmod	ECL,System
R7678	T26611	T26606	conj	System,IgG
R7679	T26612	T26610	cc	plus,ECL
R7680	T26613	T26614	compound	Western,Blotting
R7681	T26614	T26610	conj	Blotting,ECL
R7682	T26615	T26611	compound	Detection,System
R7683	T26616	T26617	punct	(,Amersham
R7684	T26617	T26611	parataxis	Amersham,System
R7685	T26618	T26617	punct	),Amersham
R7686	T26619	T26599	punct	.,detected
R7687	T27028	T27029	amod	Subcellular,analysis
R7688	T27030	T27029	compound	localization,analysis
R7689	T27032	T27033	nsubj	We,transfected
R7690	T27034	T27035	det	the,plasmid
R7691	T27035	T27033	dobj	plasmid,transfected
R7692	T27036	T27035	acl	encoding,plasmid
R7693	T27037	T27038	npadvmod	HA,tagged
R7694	T27038	T27040	amod	tagged,s
R7695	T27039	T27038	punct	-,tagged
R7696	T27040	T27036	dobj	s,encoding
R7697	T27041	T27042	amod	full,length
R7698	T27042	T27040	compound	length,s
R7699	T27043	T27042	punct	-,length
R7700	T27044	T27040	compound	mr,s
R7701	T27045	T27040	punct	-,s
R7702	T27046	T27033	prep	into,transfected
R7703	T27047	T27048	compound	HEK293T,cells
R7704	T27048	T27046	pobj	cells,into
R7705	T27049	T27033	punct	", ",transfected
R7706	T27050	T27033	conj	seeded,transfected
R7707	T27051	T27050	dobj	cells,seeded
R7708	T27052	T27050	prep	on,seeded
R7709	T27053	T27052	pobj	coverslips,on
R7710	T27054	T27053	acl	coated,coverslips
R7711	T27055	T27054	prep	with,coated
R7712	T27056	T27055	pobj	collagen,with
R7713	T27057	T27058	nummod	48,hr
R7714	T27058	T27059	npadvmod	hr,after
R7715	T27059	T27050	prep	after,seeded
R7716	T27060	T27059	pobj	transfection,after
R7717	T27061	T27050	punct	", ",seeded
R7718	T27062	T27050	conj	fixed,seeded
R7719	T27063	T27062	prep	with,fixed
R7720	T27064	T27065	nummod	4,%
R7721	T27065	T27066	compound	%,paraformaldehyde
R7722	T27066	T27063	pobj	paraformaldehyde,with
R7723	T27067	T27066	prep	in,paraformaldehyde
R7724	T27068	T27069	npadvmod	phosphate,buffered
R7725	T27069	T27071	amod	buffered,saline
R7726	T27070	T27069	punct	-,buffered
R7727	T27071	T27067	pobj	saline,in
R7728	T27072	T27071	punct	(,saline
R7729	T27073	T27071	appos	PBS,saline
R7730	T27074	T27062	punct	),fixed
R7731	T27075	T27062	prep	at,fixed
R7732	T27076	T27077	compound	room,temperature
R7733	T27077	T27075	pobj	temperature,at
R7734	T27078	T27062	prep	for,fixed
R7735	T27079	T27080	nummod	20,min
R7736	T27080	T27078	pobj	min,for
R7737	T27081	T27062	punct	", ",fixed
R7738	T27082	T27062	cc	and,fixed
R7739	T27083	T27062	conj	washed,fixed
R7740	T27084	T27083	prep	with,washed
R7741	T27085	T27084	pobj	PBS,with
R7742	T27086	T27033	punct	.,transfected
R7743	T27088	T27089	advmod	Then,permeabilized
R7744	T27090	T27089	nsubj	we,permeabilized
R7745	T27091	T27089	dobj	cells,permeabilized
R7746	T27092	T27089	prep	in,permeabilized
R7747	T27093	T27092	pobj	PBS,in
R7748	T27094	T27093	acl	containing,PBS
R7749	T27095	T27096	nummod	0.1,%
R7750	T27096	T27097	compound	%,100
R7751	T27097	T27094	dobj	100,containing
R7752	T27098	T27097	compound	Triton,100
R7753	T27099	T27097	compound	X,100
R7754	T27100	T27097	punct	-,100
R7755	T27101	T27089	prep	for,permeabilized
R7756	T27102	T27103	nummod	15,min
R7757	T27103	T27101	pobj	min,for
R7758	T27104	T27089	punct	", ",permeabilized
R7759	T27105	T27089	conj	washed,permeabilized
R7760	T27106	T27105	advmod	again,washed
R7761	T27107	T27105	prep	with,washed
R7762	T27108	T27107	pobj	PBS,with
R7763	T27109	T27105	punct	", ",washed
R7764	T27110	T27105	cc	and,washed
R7765	T27111	T27105	conj	incubated,washed
R7766	T27112	T27111	prep	in,incubated
R7767	T27113	T27112	pobj	PBS,in
R7768	T27114	T27113	acl	containing,PBS
R7769	T27115	T27116	nummod	0.1,%
R7770	T27116	T27114	dobj	%,containing
R7771	T27117	T27118	amod	bovine,albumin
R7772	T27118	T27116	appos	albumin,%
R7773	T27119	T27118	compound	serum,albumin
R7774	T27120	T27118	punct	(,albumin
R7775	T27121	T27118	appos	BSA,albumin
R7776	T27122	T27111	punct	),incubated
R7777	T27123	T27111	prep	for,incubated
R7778	T27124	T27125	nummod	30,min
R7779	T27125	T27123	pobj	min,for
R7780	T27126	T27111	prep	at,incubated
R7781	T27127	T27128	compound	room,temperature
R7782	T27128	T27126	pobj	temperature,at
R7783	T27129	T27089	punct	.,permeabilized
R7784	T27131	T27132	nsubj	We,incubated
R7785	T27133	T27132	dobj	cells,incubated
R7786	T27134	T27132	prep	with,incubated
R7787	T27135	T27136	amod	anti-HA,antibody
R7788	T27136	T27134	pobj	antibody,with
R7789	T27137	T27132	advmod	overnight,incubated
R7790	T27138	T27132	prep	at,incubated
R7791	T27139	T27140	nummod	4,°C
R7792	T27140	T27138	pobj	°C,at
R7793	T27141	T27142	punct	(,1
R7794	T27142	T27132	parataxis	1,incubated
R7795	T27143	T27144	punct	:,200
R7796	T27144	T27142	prep	200,1
R7797	T27145	T27142	prep	in,1
R7798	T27146	T27145	pobj	PBS,in
R7799	T27147	T27146	acl	containing,PBS
R7800	T27148	T27149	nummod	0.2,%
R7801	T27149	T27150	compound	%,BSA
R7802	T27150	T27147	dobj	BSA,containing
R7803	T27151	T27142	punct	),1
R7804	T27152	T27132	punct	.,incubated
R7805	T27154	T27155	det	The,day
R7806	T27155	T27157	npadvmod	day,washed
R7807	T27156	T27155	amod	following,day
R7808	T27158	T27157	punct	", ",washed
R7809	T27159	T27157	nsubj	we,washed
R7810	T27160	T27157	dobj	cells,washed
R7811	T27161	T27162	nummod	three,times
R7812	T27162	T27157	npadvmod	times,washed
R7813	T27163	T27157	prep	with,washed
R7814	T27164	T27163	pobj	PBS,with
R7815	T27165	T27157	cc	and,washed
R7816	T27166	T27157	conj	incubated,washed
R7817	T27167	T27166	prep	with,incubated
R7818	T27168	T27169	det	a,IgG
R7819	T27169	T27167	pobj	IgG,with
R7820	T27170	T27171	npadvmod	Cy3,conjugated
R7821	T27171	T27169	amod	conjugated,IgG
R7822	T27172	T27171	punct	-,conjugated
R7823	T27173	T27169	compound	goat,IgG
R7824	T27174	T27169	prep	against,IgG
R7825	T27175	T27176	compound	rabbit,IgG
R7826	T27176	T27174	pobj	IgG,against
R7827	T27177	T27178	punct	(,Laboratories
R7828	T27178	T27169	parataxis	Laboratories,IgG
R7829	T27179	T27178	dep	1,Laboratories
R7830	T27180	T27181	punct	:,400
R7831	T27181	T27179	prep	400,1
R7832	T27182	T27179	prep	in,1
R7833	T27183	T27182	pobj	PBS,in
R7834	T27184	T27178	punct	", ",Laboratories
R7835	T27185	T27178	compound	Jackson,Laboratories
R7836	T27186	T27178	compound	ImmunoReseach,Laboratories
R7837	T27187	T27178	punct	),Laboratories
R7838	T27188	T27166	prep	for,incubated
R7839	T27189	T27190	nummod	30,min
R7840	T27190	T27188	pobj	min,for
R7841	T27191	T27157	punct	.,washed
R7842	T27193	T27194	nsubj	We,rinsed
R7843	T27195	T27194	dobj	cells,rinsed
R7844	T27196	T27197	nummod	three,times
R7845	T27197	T27194	npadvmod	times,rinsed
R7846	T27198	T27194	prep	with,rinsed
R7847	T27199	T27198	pobj	PBS,with
R7848	T27200	T27194	punct	", ",rinsed
R7849	T27201	T27194	advcl	followed,rinsed
R7850	T27202	T27201	agent	by,followed
R7851	T27203	T27202	pobj	observation,by
R7852	T27204	T27203	acl	using,observation
R7853	T27205	T27206	det	a,FV300
R7854	T27206	T27204	dobj	FV300,using
R7855	T27207	T27206	amod	confocal,FV300
R7856	T27208	T27206	compound	microscope,FV300
R7857	T27209	T27210	punct	(,Olympus
R7858	T27210	T27206	parataxis	Olympus,FV300
R7859	T27211	T27210	punct	),Olympus
R7860	T27212	T27206	acl	equipped,FV300
R7861	T27213	T27212	prep	with,equipped
R7862	T27214	T27215	det	a,lens
R7863	T27215	T27213	pobj	lens,with
R7864	T27216	T27215	nummod	60,lens
R7865	T27217	T27216	punct	×,60
R7866	T27218	T27215	compound	objective,lens
R7867	T27219	T27194	punct	.,rinsed
R7870	T27909	T27910	compound	Luciferase,assay
R7871	T27912	T27913	prep	For,constructed
R7872	T27914	T27915	amod	transcriptional,analysis
R7873	T27915	T27912	pobj	analysis,For
R7874	T27916	T27915	prep	of,analysis
R7875	T27917	T27918	det	the,region
R7876	T27918	T27916	pobj	region,of
R7877	T27919	T27920	nummod	1.2,kb
R7878	T27920	T27918	compound	kb,region
R7879	T27921	T27920	punct	-,kb
R7880	T27922	T27918	compound	promoter,region
R7881	T27923	T27918	prep	of,region
R7882	T27924	T27925	compound	mr,s
R7883	T27925	T27923	pobj	s,of
R7884	T27926	T27925	punct	-,s
R7885	T27927	T27913	punct	", ",constructed
R7886	T27928	T27913	nsubj	we,constructed
R7887	T27929	T27930	det	a,plasmid
R7888	T27930	T27913	dobj	plasmid,constructed
R7889	T27931	T27930	compound	reporter,plasmid
R7890	T27932	T27933	punct	(,pro1.2k
R7891	T27933	T27930	parataxis	pro1.2k,plasmid
R7892	T27934	T27933	punct	),pro1.2k
R7893	T27935	T27913	prep	by,constructed
R7894	T27936	T27935	pcomp	subcloning,by
R7895	T27937	T27938	det	a,fragment
R7896	T27938	T27936	dobj	fragment,subcloning
R7897	T27939	T27940	nummod	1.2,kb
R7898	T27940	T27938	nmod	kb,fragment
R7899	T27941	T27940	punct	-,kb
R7900	T27942	T27938	amod	upstream,fragment
R7901	T27943	T27938	amod	genomic,fragment
R7902	T27944	T27938	prep	of,fragment
R7903	T27945	T27946	compound	mr,s
R7904	T27946	T27948	compound	s,gene
R7905	T27947	T27946	punct	-,s
R7906	T27948	T27944	pobj	gene,of
R7907	T27949	T27936	prep	into,subcloning
R7908	T27950	T27951	det	a,plasmid
R7909	T27951	T27949	pobj	plasmid,into
R7910	T27952	T27953	compound	pGL3,reporter
R7911	T27953	T27951	compound	reporter,plasmid
R7912	T27954	T27953	compound	luciferase,reporter
R7913	T27955	T27956	punct	(,Promega
R7914	T27956	T27951	parataxis	Promega,plasmid
R7915	T27957	T27956	punct	),Promega
R7916	T27958	T27913	punct	.,constructed
R7917	T27960	T27961	det	The,vectors
R7918	T27961	T27963	nsubjpass	vectors,constructed
R7919	T27962	T27961	compound	expression,vectors
R7920	T27964	T27963	auxpass	were,constructed
R7921	T27965	T27963	prep	by,constructed
R7922	T27966	T27965	pcomp	subcloning,by
R7923	T27967	T27968	amod	full,length
R7924	T27968	T27970	nmod	length,genes
R7925	T27969	T27968	punct	-,length
R7926	T27970	T27966	dobj	genes,subcloning
R7927	T27971	T27972	nmod	Crx,Nrl
R7928	T27972	T27970	compound	Nrl,genes
R7929	T27973	T27972	punct	", ",Nrl
R7930	T27974	T27972	nmod	Otx2,Nrl
R7931	T27975	T27972	punct	", ",Nrl
R7932	T27976	T27966	prep	into,subcloning
R7933	T27977	T27978	det	a,vector
R7934	T27978	T27976	pobj	vector,into
R7935	T27979	T27978	compound	pMIK,vector
R7936	T27980	T27978	compound	expression,vector
R7937	T27981	T27982	punct	(,gift
R7938	T27982	T27978	parataxis	gift,vector
R7939	T27983	T27982	det	a,gift
R7940	T27984	T27982	prep	from,gift
R7941	T27985	T27986	compound	Dr.,Maruyama
R7942	T27986	T27984	pobj	Maruyama,from
R7943	T27987	T27986	compound	K.,Maruyama
R7944	T27988	T27982	punct	),gift
R7945	T27989	T27966	punct	", ",subcloning
R7946	T27990	T27966	advmod	respectively,subcloning
R7947	T27991	T27963	punct	.,constructed
R7948	T27993	T27994	prep	For,mutated
R7949	T27995	T27996	det	the,vector
R7950	T27996	T27993	pobj	vector,For
R7951	T27997	T27996	punct	"""",vector
R7952	T27998	T27996	nmod	mut1259,vector
R7953	T27999	T27996	punct	"""",vector
R7954	T28000	T27994	punct	", ",mutated
R7955	T28001	T28002	det	the,site
R7956	T28002	T27994	nsubjpass	site,mutated
R7957	T28003	T28004	compound	Crx,binding
R7958	T28004	T28002	compound	binding,site
R7959	T28005	T28002	prep	at,site
R7960	T28006	T28007	det	the,position
R7961	T28007	T28005	pobj	position,at
R7962	T28008	T28009	punct	-,1259
R7963	T28009	T28010	nummod	1259,bp
R7964	T28010	T28007	compound	bp,position
R7965	T28011	T27994	auxpass	was,mutated
R7966	T28012	T27994	prep	by,mutated
R7967	T28013	T28012	pcomp	replacing,by
R7968	T28014	T28013	dobj	GGATTA,replacing
R7969	T28015	T28013	prep	with,replacing
R7970	T28016	T28015	pobj	AGATCT,with
R7971	T28017	T27994	punct	.,mutated
R7972	T28019	T28020	prep	For,mutated
R7973	T28021	T28022	det	the,vector
R7974	T28022	T28019	pobj	vector,For
R7975	T28023	T28022	punct	"""",vector
R7976	T28024	T28022	nmod	mut198,vector
R7977	T28025	T28022	punct	"""",vector
R7978	T28026	T28020	punct	", ",mutated
R7979	T28027	T28028	det	the,site
R7980	T28028	T28020	nsubjpass	site,mutated
R7981	T28029	T28030	compound	Crx,binding
R7982	T28030	T28028	compound	binding,site
R7983	T28031	T28028	prep	at,site
R7984	T28032	T28033	det	the,position
R7985	T28033	T28031	pobj	position,at
R7986	T28034	T28035	punct	-,198
R7987	T28035	T28036	nummod	198,bp
R7988	T28036	T28033	compound	bp,position
R7989	T28037	T28020	auxpass	was,mutated
R7990	T28038	T28020	prep	by,mutated
R7991	T28039	T28038	pcomp	replacing,by
R7992	T28040	T28039	dobj	TAATCC,replacing
R7993	T28041	T28039	prep	with,replacing
R7994	T28042	T28041	pobj	GAATTC,with
R7995	T28043	T28020	punct	.,mutated
R7996	T28045	T28046	prep	For,mutated
R7997	T28047	T28048	det	the,vector
R7998	T28048	T28045	pobj	vector,For
R7999	T28049	T28048	punct	"""",vector
R8000	T28050	T28048	nmod	mut72,vector
R8001	T28051	T28048	punct	"""",vector
R8002	T28052	T28046	punct	", ",mutated
R8003	T28053	T28054	det	the,site
R8004	T28054	T28046	nsubjpass	site,mutated
R8005	T28055	T28056	compound	Crx,binding
R8006	T28056	T28054	compound	binding,site
R8007	T28057	T28054	prep	at,site
R8008	T28058	T28059	det	the,position
R8009	T28059	T28057	pobj	position,at
R8010	T28060	T28061	punct	-,72
R8011	T28061	T28062	nummod	72,bp
R8012	T28062	T28059	compound	bp,position
R8013	T28063	T28046	auxpass	was,mutated
R8014	T28064	T28046	prep	by,mutated
R8015	T28065	T28064	pcomp	replacing,by
R8016	T28066	T28065	dobj	GGATTA,replacing
R8017	T28067	T28065	prep	with,replacing
R8018	T28068	T28067	pobj	GAATTC,with
R8019	T28069	T28046	punct	.,mutated
R8020	T28071	T28072	prep	For,mutated
R8021	T28073	T28074	det	the,vector
R8022	T28074	T28071	pobj	vector,For
R8023	T28075	T28074	punct	"""",vector
R8024	T28076	T28077	nmod	mut,all
R8025	T28077	T28074	nmod	all,vector
R8026	T28078	T28074	punct	"""",vector
R8027	T28079	T28072	punct	", ",mutated
R8028	T28080	T28072	nsubjpass	all,mutated
R8029	T28081	T28080	prep	of,all
R8030	T28082	T28083	nummod	three,sites
R8031	T28083	T28081	pobj	sites,of
R8032	T28084	T28085	compound	Crx,binding
R8033	T28085	T28083	compound	binding,sites
R8034	T28086	T28072	auxpass	were,mutated
R8035	T28087	T28072	punct	.,mutated
R8036	T28089	T28090	compound	5xGAL4,pGL3
R8037	T28090	T28092	compound	pGL3,plasmid
R8038	T28091	T28090	punct	-,pGL3
R8039	T28092	T28095	nsubjpass	plasmid,gifted
R8040	T28093	T28092	compound	Control,plasmid
R8041	T28094	T28092	compound	reporter,plasmid
R8042	T28096	T28092	punct	", ",plasmid
R8043	T28097	T28098	dep	which,contains
R8044	T28098	T28092	relcl	contains,plasmid
R8045	T28099	T28100	nummod	five,copies
R8046	T28100	T28098	dobj	copies,contains
R8047	T28101	T28100	prep	of,copies
R8048	T28102	T28103	det	the,sequence
R8049	T28103	T28101	pobj	sequence,of
R8050	T28104	T28103	compound	GAL4,sequence
R8051	T28105	T28106	compound	DNA,recognition
R8052	T28106	T28103	compound	recognition,sequence
R8053	T28107	T28100	acl	positioned,copies
R8054	T28108	T28107	advmod	immediately,positioned
R8055	T28109	T28108	advmod	upstream,immediately
R8056	T28110	T28108	prep	of,immediately
R8057	T28111	T28112	nmod	SV40,promoter
R8058	T28112	T28110	pobj	promoter,of
R8059	T28113	T28112	amod	minimal,promoter
R8060	T28114	T28095	punct	", ",gifted
R8061	T28115	T28095	auxpass	was,gifted
R8062	T28116	T28095	advmod	kindly,gifted
R8063	T28117	T28095	agent	from,gifted
R8064	T28118	T28119	compound	Dr.,Noguchi
R8065	T28119	T28117	pobj	Noguchi,from
R8066	T28120	T28119	compound	T.,Noguchi
R8067	T28121	T28095	cc	and,gifted
R8068	T28122	T28095	conj	used,gifted
R8069	T28123	T28122	prep	for,used
R8070	T28124	T28123	pobj	analysis,for
R8071	T28125	T28124	prep	of,analysis
R8072	T28126	T28127	det	the,activity
R8073	T28127	T28125	pobj	activity,of
R8074	T28128	T28127	amod	transcriptional,activity
R8075	T28129	T28127	prep	of,activity
R8076	T28130	T28131	compound	mr,s
R8077	T28131	T28129	pobj	s,of
R8078	T28132	T28131	punct	-,s
R8079	T28133	T28134	punct	[,50
R8080	T28134	T28122	parataxis	50,used
R8081	T28135	T28134	punct	],50
R8082	T28136	T28095	punct	.,gifted
R8083	T28138	T28139	aux	To,generate
R8084	T28139	T28140	advcl	generate,produced
R8085	T28141	T28142	compound	effector,plasmids
R8086	T28142	T28139	dobj	plasmids,generate
R8087	T28143	T28140	punct	", ",produced
R8088	T28144	T28145	amod	various,fragments
R8089	T28145	T28140	nsubj	fragments,produced
R8090	T28146	T28145	compound	deletion,fragments
R8091	T28147	T28140	aux	were,produced
R8092	T28148	T28140	prep	by,produced
R8093	T28149	T28150	compound	PCR,reaction
R8094	T28150	T28148	pobj	reaction,by
R8095	T28151	T28140	cc	and,produced
R8096	T28152	T28140	conj	subcloned,produced
R8097	T28153	T28152	prep	into,subcloned
R8098	T28154	T28155	compound	pGBKT7,vector
R8099	T28155	T28153	pobj	vector,into
R8100	T28156	T28140	punct	", ",produced
R8101	T28157	T28140	advmod	respectively,produced
R8102	T28158	T28140	punct	.,produced
R8103	T28160	T28161	det	These,fragments
R8104	T28161	T28162	nsubjpass	fragments,digested
R8105	T28163	T28162	auxpass	were,digested
R8106	T28164	T28162	prep	with,digested
R8107	T28165	T28166	det	the,sequence
R8108	T28166	T28164	pobj	sequence,with
R8109	T28167	T28166	punct	", ",sequence
R8110	T28168	T28169	dep	which,encodes
R8111	T28169	T28166	relcl	encodes,sequence
R8112	T28170	T28171	compound	GAL4,domain
R8113	T28171	T28169	dobj	domain,encodes
R8114	T28172	T28173	compound	DNA,binding
R8115	T28173	T28171	compound	binding,domain
R8116	T28174	T28162	punct	", ",digested
R8117	T28175	T28162	cc	and,digested
R8118	T28176	T28162	conj	inserted,digested
R8119	T28177	T28176	prep	into,inserted
R8120	T28178	T28177	pobj	pcDNA3,into
R8121	T28179	T28162	punct	.,digested
R8122	T28181	T28182	nsubj	We,transfected
R8123	T28183	T28184	nummod	0.1,μg
R8124	T28184	T28182	dobj	μg,transfected
R8125	T28185	T28184	prep	of,μg
R8126	T28186	T28187	compound	reporter,DNA
R8127	T28187	T28185	pobj	DNA,of
R8128	T28188	T28187	compound	plasmid,DNA
R8129	T28189	T28184	cc	and,μg
R8130	T28190	T28191	nummod	2,μg
R8131	T28191	T28184	conj	μg,μg
R8132	T28192	T28191	prep	of,μg
R8133	T28193	T28194	det	the,DNA
R8134	T28194	T28192	pobj	DNA,of
R8135	T28195	T28196	compound	expression,vector
R8136	T28196	T28194	compound	vector,DNA
R8137	T28197	T28184	prep	per,μg
R8138	T28198	T28199	nummod	6,cm
R8139	T28199	T28200	compound	cm,dish
R8140	T28200	T28197	pobj	dish,per
R8141	T28201	T28182	prep	into,transfected
R8142	T28202	T28203	compound	HEK293T,cells
R8143	T28203	T28201	pobj	cells,into
R8144	T28204	T28182	advcl	using,transfected
R8145	T28205	T28206	compound	Fugene6,reagent
R8146	T28206	T28204	dobj	reagent,using
R8147	T28207	T28206	compound	transfection,reagent
R8148	T28208	T28182	punct	.,transfected
R8149	T28210	T28211	nsubj	We,analyzed
R8150	T28212	T28213	compound	luciferase,activity
R8151	T28213	T28211	dobj	activity,analyzed
R8152	T28214	T28215	nummod	48,hr
R8153	T28215	T28216	npadvmod	hr,after
R8154	T28216	T28211	prep	after,analyzed
R8155	T28217	T28216	pobj	transfection,after
R8156	T28218	T28211	punct	.,analyzed

PMC:1435744 / 40427-48657 JSONTXT 4 Projects

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PMC:1435744 / 40427-48657 JSON TXT 4 Projects