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PMC:1142324 / 16149-24003 JSONTXT 4 Projects

Annnotations TAB TSV DIC JSON TextAE

Id Subject Object Predicate Lexical cue
T4651 9-21 NN denotes Construction
T4652 22-24 IN denotes of
T4653 25-27 DT denotes an
T4655 28-34 NN denotes Adam22
T4656 35-39 NN denotes gene
T4658 39-40 HYPH denotes -
T4657 40-49 VBG denotes targeting
T4654 50-56 NN denotes vector
T4659 56-278 sentence denotes The 18-kb genomic DNA fragments covering exons 5, 6, 7, 8 and 9 of the Adam22 gene were amplified from C57BL/6 genomic DNA by PCR using Expand Hi-Fidelity enzyme mix (Roche Diagnostics) and primers designed for each exon.
T4660 57-60 DT denotes The
T4662 61-63 CD denotes 18
T4664 63-64 HYPH denotes -
T4663 64-66 NN denotes kb
T4665 67-74 JJ denotes genomic
T4666 75-78 NN denotes DNA
T4661 79-88 NNS denotes fragments
T4668 89-97 VBG denotes covering
T4669 98-103 NNS denotes exons
T4670 104-105 CD denotes 5
T4671 105-107 , denotes ,
T4672 107-108 CD denotes 6
T4673 108-110 , denotes ,
T4674 110-111 CD denotes 7
T4675 111-113 , denotes ,
T4676 113-114 CD denotes 8
T4677 115-118 CC denotes and
T4678 119-120 CD denotes 9
T4679 121-123 IN denotes of
T4680 124-127 DT denotes the
T4682 128-134 NN denotes Adam22
T4681 135-139 NN denotes gene
T4683 140-144 VBD denotes were
T4667 145-154 VBN denotes amplified
T4684 155-159 IN denotes from
T4685 160-165 NN denotes C57BL
T4687 165-166 HYPH denotes /
T4688 166-167 CD denotes 6
T4689 168-175 JJ denotes genomic
T4686 176-179 NN denotes DNA
T4690 180-182 IN denotes by
T4691 183-186 NN denotes PCR
T4692 187-192 VBG denotes using
T4693 193-199 NNP denotes Expand
T4695 200-202 NNP denotes Hi
T4697 202-203 HYPH denotes -
T4696 203-211 NNP denotes Fidelity
T4698 212-218 NN denotes enzyme
T4694 219-222 NN denotes mix
T4699 223-224 -LRB- denotes (
T4701 224-229 NNP denotes Roche
T4700 230-241 NNP denotes Diagnostics
T4702 241-242 -RRB- denotes )
T4703 243-246 CC denotes and
T4704 247-254 NNS denotes primers
T4705 255-263 VBN denotes designed
T4706 264-267 IN denotes for
T4707 268-272 DT denotes each
T4708 273-277 NN denotes exon
T4709 277-278 . denotes .
T4710 278-391 sentence denotes To generate a mutation in the mouse Adam22 gene, we inserted the PGKneo cassette into exon 8 of the Adam22 gene.
T4711 279-281 TO denotes To
T4712 282-290 VB denotes generate
T4714 291-292 DT denotes a
T4715 293-301 NN denotes mutation
T4716 302-304 IN denotes in
T4717 305-308 DT denotes the
T4719 309-314 NN denotes mouse
T4720 315-321 NN denotes Adam22
T4718 322-326 NN denotes gene
T4721 326-328 , denotes ,
T4722 328-330 PRP denotes we
T4713 331-339 VBD denotes inserted
T4723 340-343 DT denotes the
T4725 344-350 NN denotes PGKneo
T4724 351-359 NN denotes cassette
T4726 360-364 IN denotes into
T4727 365-369 NN denotes exon
T4728 370-371 CD denotes 8
T4729 372-374 IN denotes of
T4730 375-378 DT denotes the
T4732 379-385 NN denotes Adam22
T4731 386-390 NN denotes gene
T4733 390-391 . denotes .
T4734 391-621 sentence denotes The final targeting construct consisted of a 10.5-kb genomic DNA fragment that was interrupted by the insertion of the PGKneo cassette and contained a MC1/TK cassette as a negative selection marker against random integration [5].
T4735 392-395 DT denotes The
T4737 396-401 JJ denotes final
T4738 402-411 NN denotes targeting
T4736 412-421 NN denotes construct
T4739 422-431 VBD denotes consisted
T4740 432-434 IN denotes of
T4741 435-436 DT denotes a
T4743 437-441 CD denotes 10.5
T4745 441-442 HYPH denotes -
T4744 442-444 NN denotes kb
T4746 445-452 JJ denotes genomic
T4747 453-456 NN denotes DNA
T4742 457-465 NN denotes fragment
T4748 466-470 WDT denotes that
T4750 471-474 VBD denotes was
T4749 475-486 VBN denotes interrupted
T4751 487-489 IN denotes by
T4752 490-493 DT denotes the
T4753 494-503 NN denotes insertion
T4754 504-506 IN denotes of
T4755 507-510 DT denotes the
T4757 511-517 NN denotes PGKneo
T4756 518-526 NN denotes cassette
T4758 527-530 CC denotes and
T4759 531-540 VBD denotes contained
T4760 541-542 DT denotes a
T4762 543-546 NN denotes MC1
T4764 546-547 HYPH denotes /
T4763 547-549 NN denotes TK
T4761 550-558 NN denotes cassette
T4765 559-561 IN denotes as
T4766 562-563 DT denotes a
T4768 564-572 JJ denotes negative
T4769 573-582 NN denotes selection
T4767 583-589 NN denotes marker
T4770 590-597 IN denotes against
T4771 598-604 JJ denotes random
T4772 605-616 NN denotes integration
T4773 617-618 -LRB- denotes [
T4774 618-619 CD denotes 5
T4775 619-620 -RRB- denotes ]
T4776 620-621 . denotes .
T4851 623-633 NN denotes Generation
T4852 634-636 IN denotes of
T4853 637-643 NN denotes Adam22
T4854 644-653 JJ denotes deficient
T4855 654-658 NNS denotes mice
T4856 658-750 sentence denotes The linearised targeting vector was electroporated into TT2 embryonic stem (ES) cells [31].
T4857 659-662 DT denotes The
T4859 663-673 VBN denotes linearised
T4860 674-683 NN denotes targeting
T4858 684-690 NN denotes vector
T4862 691-694 VBD denotes was
T4861 695-709 VBN denotes electroporated
T4863 710-714 IN denotes into
T4864 715-718 NN denotes TT2
T4866 719-728 JJ denotes embryonic
T4867 729-733 NN denotes stem
T4868 734-735 -LRB- denotes (
T4869 735-737 NN denotes ES
T4870 737-738 -RRB- denotes )
T4865 739-744 NNS denotes cells
T4871 745-746 -LRB- denotes [
T4872 746-748 CD denotes 31
T4873 748-749 -RRB- denotes ]
T4874 749-750 . denotes .
T4875 750-796 sentence denotes Homologous recombinants were selected by PCR.
T4876 751-761 JJ denotes Homologous
T4877 762-774 NNS denotes recombinants
T4879 775-779 VBD denotes were
T4878 780-788 VBN denotes selected
T4880 789-791 IN denotes by
T4881 792-795 NN denotes PCR
T4882 795-796 . denotes .
T4883 796-1041 sentence denotes The extracted genomic DNA from each clone was amplified using the forward primer AGN2: 5'-GCCTGCTTGCCGAATATCATGGTGGAAAAT-3' in the PGKneo cassette, and the reverse primer MFP065R: 5'-ACTATTTCTGTGATGAGGGCACAGCATC-3' outside the targeting vector.
T4884 797-800 DT denotes The
T4886 801-810 VBN denotes extracted
T4887 811-818 JJ denotes genomic
T4885 819-822 NN denotes DNA
T4889 823-827 IN denotes from
T4890 828-832 DT denotes each
T4891 833-838 NN denotes clone
T4892 839-842 VBD denotes was
T4888 843-852 VBN denotes amplified
T4893 853-858 VBG denotes using
T4894 859-862 DT denotes the
T4896 863-870 JJ denotes forward
T4895 871-877 NN denotes primer
T4897 878-882 NN denotes AGN2
T4898 882-884 : denotes :
T4899 884-885 CD denotes 5
T4901 885-886 SYM denotes '
T4902 886-887 HYPH denotes -
T4900 887-917 NN denotes GCCTGCTTGCCGAATATCATGGTGGAAAAT
T4903 917-918 HYPH denotes -
T4904 918-919 CD denotes 3
T4905 919-920 SYM denotes '
T4906 921-923 IN denotes in
T4907 924-927 DT denotes the
T4909 928-934 NN denotes PGKneo
T4908 935-943 NN denotes cassette
T4910 943-945 , denotes ,
T4911 945-948 CC denotes and
T4912 949-952 DT denotes the
T4914 953-960 JJ denotes reverse
T4913 961-967 NN denotes primer
T4915 968-975 NN denotes MFP065R
T4916 975-977 : denotes :
T4917 977-978 CD denotes 5
T4919 978-979 SYM denotes '
T4920 979-980 HYPH denotes -
T4918 980-1008 NN denotes ACTATTTCTGTGATGAGGGCACAGCATC
T4921 1008-1009 HYPH denotes -
T4922 1009-1010 CD denotes 3
T4923 1010-1011 SYM denotes '
T4924 1012-1019 IN denotes outside
T4925 1020-1023 DT denotes the
T4927 1024-1033 NN denotes targeting
T4926 1034-1040 NN denotes vector
T4928 1040-1041 . denotes .
T4929 1041-1171 sentence denotes Homologous recombined DNA was efficiently amplified by 35 cycles of the following amplification steps (94°C-30 s and 68°C-5 min).
T4930 1042-1052 JJ denotes Homologous
T4932 1053-1063 JJ denotes recombined
T4931 1064-1067 NN denotes DNA
T4934 1068-1071 VBD denotes was
T4935 1072-1083 RB denotes efficiently
T4933 1084-1093 VBN denotes amplified
T4936 1094-1096 IN denotes by
T4937 1097-1099 CD denotes 35
T4938 1100-1106 NNS denotes cycles
T4939 1107-1109 IN denotes of
T4940 1110-1113 DT denotes the
T4942 1114-1123 VBG denotes following
T4943 1124-1137 NN denotes amplification
T4941 1138-1143 NNS denotes steps
T4944 1144-1145 -LRB- denotes (
T4945 1145-1147 CD denotes 94
T4946 1147-1149 NN denotes °C
T4947 1149-1150 HYPH denotes -
T4948 1150-1152 CD denotes 30
T4949 1153-1154 NN denotes s
T4950 1155-1158 CC denotes and
T4951 1159-1161 CD denotes 68
T4952 1161-1163 NN denotes °C
T4953 1163-1164 HYPH denotes -
T4954 1164-1165 CD denotes 5
T4955 1166-1169 NN denotes min
T4956 1169-1170 -RRB- denotes )
T4957 1170-1171 . denotes .
T4958 1171-1228 sentence denotes The targeting efficiency of this construct was about 4%.
T4959 1172-1175 DT denotes The
T4961 1176-1185 NN denotes targeting
T4960 1186-1196 NN denotes efficiency
T4963 1197-1199 IN denotes of
T4964 1200-1204 DT denotes this
T4965 1205-1214 NN denotes construct
T4962 1215-1218 VBD denotes was
T4966 1219-1224 RB denotes about
T4967 1225-1226 CD denotes 4
T4968 1226-1227 NN denotes %
T4969 1227-1228 . denotes .
T4970 1228-1327 sentence denotes Correctly targeted ES clones were injected into fertilized ICR mouse eggs at the eight-cell stage.
T4971 1229-1238 RB denotes Correctly
T4972 1239-1247 VBN denotes targeted
T4974 1248-1250 NN denotes ES
T4973 1251-1257 NNS denotes clones
T4976 1258-1262 VBD denotes were
T4975 1263-1271 VBN denotes injected
T4977 1272-1276 IN denotes into
T4978 1277-1287 VBN denotes fertilized
T4980 1288-1291 NN denotes ICR
T4981 1292-1297 NN denotes mouse
T4979 1298-1302 NNS denotes eggs
T4982 1303-1305 IN denotes at
T4983 1306-1309 DT denotes the
T4985 1310-1315 CD denotes eight
T4987 1315-1316 HYPH denotes -
T4986 1316-1320 NN denotes cell
T4984 1321-1326 NN denotes stage
T4988 1326-1327 . denotes .
T4989 1327-1471 sentence denotes The resulting male chimeras were mated with C57BL/6N females, and heterozygous male and female mice were interbred to generate homozygous mice.
T4990 1328-1331 DT denotes The
T4992 1332-1341 VBG denotes resulting
T4993 1342-1346 JJ denotes male
T4991 1347-1355 NNS denotes chimeras
T4995 1356-1360 VBD denotes were
T4994 1361-1366 VBN denotes mated
T4996 1367-1371 IN denotes with
T4997 1372-1377 NN denotes C57BL
T4999 1377-1378 HYPH denotes /
T5000 1378-1380 CD denotes 6N
T4998 1381-1388 NNS denotes females
T5001 1388-1390 , denotes ,
T5002 1390-1393 CC denotes and
T5003 1394-1406 JJ denotes heterozygous
T5005 1407-1411 JJ denotes male
T5006 1412-1415 CC denotes and
T5007 1416-1422 JJ denotes female
T5004 1423-1427 NNS denotes mice
T5009 1428-1432 VBD denotes were
T5008 1433-1442 VBN denotes interbred
T5010 1443-1445 TO denotes to
T5011 1446-1454 VB denotes generate
T5012 1455-1465 JJ denotes homozygous
T5013 1466-1470 NNS denotes mice
T5014 1470-1471 . denotes .
T5015 1471-1552 sentence denotes The ataxic phenotypes of homozygous mice were observed in two independent lines.
T5016 1472-1475 DT denotes The
T5018 1476-1482 JJ denotes ataxic
T5017 1483-1493 NNS denotes phenotypes
T5020 1494-1496 IN denotes of
T5021 1497-1507 JJ denotes homozygous
T5022 1508-1512 NNS denotes mice
T5023 1513-1517 VBD denotes were
T5019 1518-1526 VBN denotes observed
T5024 1527-1529 IN denotes in
T5025 1530-1533 CD denotes two
T5027 1534-1545 JJ denotes independent
T5026 1546-1551 NNS denotes lines
T5028 1551-1552 . denotes .
T5055 1554-1562 NNP denotes Southern
T5056 1563-1567 NN denotes blot
T5057 1568-1576 NN denotes analysis
T5058 1576-1673 sentence denotes Mouse genomic DNA used for Southern blot analysis was extracted from the liver of each genotype.
T5059 1577-1582 NN denotes Mouse
T5061 1583-1590 JJ denotes genomic
T5060 1591-1594 NN denotes DNA
T5063 1595-1599 VBN denotes used
T5064 1600-1603 IN denotes for
T5065 1604-1612 NNP denotes Southern
T5066 1613-1617 NN denotes blot
T5067 1618-1626 NN denotes analysis
T5068 1627-1630 VBD denotes was
T5062 1631-1640 VBN denotes extracted
T5069 1641-1645 IN denotes from
T5070 1646-1649 DT denotes the
T5071 1650-1655 NN denotes liver
T5072 1656-1658 IN denotes of
T5073 1659-1663 DT denotes each
T5074 1664-1672 NN denotes genotype
T5075 1672-1673 . denotes .
T5076 1673-1811 sentence denotes BamHI-digested genomic DNA was probed with the [32P]-labelled 0.4-kb SpeI-BamHI genomic fragment, which is located between exons 8 and 9.
T5077 1674-1679 NN denotes BamHI
T5079 1679-1680 HYPH denotes -
T5078 1680-1688 VBN denotes digested
T5081 1689-1696 JJ denotes genomic
T5080 1697-1700 NN denotes DNA
T5083 1701-1704 VBD denotes was
T5082 1705-1711 VBN denotes probed
T5084 1712-1716 IN denotes with
T5085 1717-1720 DT denotes the
T5087 1721-1722 -LRB- denotes [
T5088 1722-1725 CD denotes 32P
T5090 1725-1726 -RRB- denotes ]
T5091 1726-1727 HYPH denotes -
T5089 1727-1735 VBN denotes labelled
T5092 1736-1739 CD denotes 0.4
T5094 1739-1740 HYPH denotes -
T5093 1740-1742 NN denotes kb
T5095 1743-1747 NN denotes SpeI
T5097 1747-1748 HYPH denotes -
T5096 1748-1753 NN denotes BamHI
T5098 1754-1761 JJ denotes genomic
T5086 1762-1770 NN denotes fragment
T5099 1770-1772 , denotes ,
T5100 1772-1777 WDT denotes which
T5102 1778-1780 VBZ denotes is
T5101 1781-1788 VBN denotes located
T5103 1789-1796 IN denotes between
T5104 1797-1802 NNS denotes exons
T5105 1803-1804 CD denotes 8
T5106 1805-1808 CC denotes and
T5107 1809-1810 CD denotes 9
T5108 1810-1811 . denotes .
T5223 1813-1821 NN denotes Antibody
T5224 1822-1832 NN denotes production
T5225 1832-2011 sentence denotes His-tagged and MBP-fused recombinant protein containing cytoplasmic domain (cp) of human ADAM22 isoform 1 (position: 756–906, 151 amino acids in length) were produced in E. coli.
T5226 1833-1836 NN denotes His
T5228 1836-1837 HYPH denotes -
T5227 1837-1843 VBN denotes tagged
T5230 1844-1847 CC denotes and
T5231 1848-1851 NN denotes MBP
T5233 1851-1852 HYPH denotes -
T5232 1852-1857 VBN denotes fused
T5234 1858-1869 JJ denotes recombinant
T5229 1870-1877 NN denotes protein
T5236 1878-1888 VBG denotes containing
T5237 1889-1900 JJ denotes cytoplasmic
T5238 1901-1907 NN denotes domain
T5239 1908-1909 -LRB- denotes (
T5240 1909-1911 NN denotes cp
T5241 1911-1912 -RRB- denotes )
T5242 1913-1915 IN denotes of
T5243 1916-1921 JJ denotes human
T5245 1922-1928 NN denotes ADAM22
T5244 1929-1936 NN denotes isoform
T5246 1937-1938 CD denotes 1
T5247 1939-1940 -LRB- denotes (
T5249 1940-1948 NN denotes position
T5250 1948-1950 : denotes :
T5251 1950-1953 CD denotes 756
T5253 1953-1954 SYM denotes –
T5252 1954-1957 CD denotes 906
T5254 1957-1959 , denotes ,
T5255 1959-1962 CD denotes 151
T5256 1963-1968 NN denotes amino
T5248 1969-1974 NNS denotes acids
T5257 1975-1977 IN denotes in
T5258 1978-1984 NN denotes length
T5259 1984-1985 -RRB- denotes )
T5260 1986-1990 VBD denotes were
T5235 1991-1999 VBN denotes produced
T5261 2000-2002 IN denotes in
T5262 2003-2005 NNP denotes E.
T5263 2006-2010 NNP denotes coli
T5264 2010-2011 . denotes .
T5265 2011-2130 sentence denotes His-tagged ADAM22-cp protein was purified in denatured condition using Ni-NTA resin (Qiagen GmbH) and dialysed in PBS.
T5266 2012-2015 NN denotes His
T5268 2015-2016 HYPH denotes -
T5267 2016-2022 VBN denotes tagged
T5270 2023-2029 NN denotes ADAM22
T5272 2029-2030 HYPH denotes -
T5271 2030-2032 NN denotes cp
T5269 2033-2040 NN denotes protein
T5274 2041-2044 VBD denotes was
T5273 2045-2053 VBN denotes purified
T5275 2054-2056 IN denotes in
T5276 2057-2066 VBN denotes denatured
T5277 2067-2076 NN denotes condition
T5278 2077-2082 VBG denotes using
T5279 2083-2085 NN denotes Ni
T5281 2085-2086 HYPH denotes -
T5280 2086-2089 NN denotes NTA
T5282 2090-2095 NN denotes resin
T5283 2096-2097 -LRB- denotes (
T5285 2097-2103 NNP denotes Qiagen
T5284 2104-2108 NNP denotes GmbH
T5286 2108-2109 -RRB- denotes )
T5287 2110-2113 CC denotes and
T5288 2114-2122 VBN denotes dialysed
T5289 2123-2125 IN denotes in
T5290 2126-2129 NN denotes PBS
T5291 2129-2130 . denotes .
T5292 2130-2281 sentence denotes Precipitated protein was recovered and was mixed with Freund's complete adjuvant (Invitrogen), then, the mixture was used for immunization of rabbits.
T5293 2131-2143 VBN denotes Precipitated
T5294 2144-2151 NN denotes protein
T5296 2152-2155 VBD denotes was
T5295 2156-2165 VBN denotes recovered
T5298 2166-2169 CC denotes and
T5299 2170-2173 VBD denotes was
T5300 2174-2179 VBN denotes mixed
T5301 2180-2184 IN denotes with
T5302 2185-2191 NNP denotes Freund
T5304 2191-2193 POS denotes 's
T5305 2194-2202 JJ denotes complete
T5303 2203-2211 NN denotes adjuvant
T5306 2212-2213 -LRB- denotes (
T5307 2213-2223 NNP denotes Invitrogen
T5308 2223-2224 -RRB- denotes )
T5309 2224-2226 , denotes ,
T5310 2226-2230 RB denotes then
T5311 2230-2232 , denotes ,
T5312 2232-2235 DT denotes the
T5313 2236-2243 NN denotes mixture
T5314 2244-2247 VBD denotes was
T5297 2248-2252 VBN denotes used
T5315 2253-2256 IN denotes for
T5316 2257-2269 NN denotes immunization
T5317 2270-2272 IN denotes of
T5318 2273-2280 NNS denotes rabbits
T5319 2280-2281 . denotes .
T5320 2281-2430 sentence denotes The antiserum raised against the His-tagged ADAM22-cp protein was incubated with MBP-fused ADAM22-cp protein coupled to Affi-Gel 10 beads (Bio-Rad).
T5321 2282-2285 DT denotes The
T5322 2286-2295 NN denotes antiserum
T5324 2296-2302 VBN denotes raised
T5325 2303-2310 IN denotes against
T5326 2311-2314 DT denotes the
T5328 2315-2318 NN denotes His
T5330 2318-2319 HYPH denotes -
T5329 2319-2325 VBN denotes tagged
T5331 2326-2332 NN denotes ADAM22
T5333 2332-2333 HYPH denotes -
T5332 2333-2335 NN denotes cp
T5327 2336-2343 NN denotes protein
T5334 2344-2347 VBD denotes was
T5323 2348-2357 VBN denotes incubated
T5335 2358-2362 IN denotes with
T5336 2363-2366 NN denotes MBP
T5338 2366-2367 HYPH denotes -
T5337 2367-2372 VBN denotes fused
T5340 2373-2379 NN denotes ADAM22
T5342 2379-2380 HYPH denotes -
T5341 2380-2382 NN denotes cp
T5339 2383-2390 NN denotes protein
T5343 2391-2398 VBN denotes coupled
T5344 2399-2401 IN denotes to
T5345 2402-2406 NN denotes Affi
T5347 2406-2407 HYPH denotes -
T5346 2407-2410 NN denotes Gel
T5349 2411-2413 CD denotes 10
T5348 2414-2419 NNS denotes beads
T5350 2420-2421 -LRB- denotes (
T5352 2421-2424 NNP denotes Bio
T5353 2424-2425 HYPH denotes -
T5351 2425-2428 NNP denotes Rad
T5354 2428-2429 -RRB- denotes )
T5355 2429-2430 . denotes .
T5356 2430-2573 sentence denotes Beads with bound antibodies were washed in PBS, and the bound antibodies were eluted with 100 mM glycine, pH 3.0, and neutralized immediately.
T5357 2431-2436 NNS denotes Beads
T5359 2437-2441 IN denotes with
T5360 2442-2447 VBN denotes bound
T5361 2448-2458 NNS denotes antibodies
T5362 2459-2463 VBD denotes were
T5358 2464-2470 VBN denotes washed
T5363 2471-2473 IN denotes in
T5364 2474-2477 NN denotes PBS
T5365 2477-2479 , denotes ,
T5366 2479-2482 CC denotes and
T5367 2483-2486 DT denotes the
T5369 2487-2492 VBN denotes bound
T5368 2493-2503 NNS denotes antibodies
T5371 2504-2508 VBD denotes were
T5370 2509-2515 VBN denotes eluted
T5372 2516-2520 IN denotes with
T5373 2521-2524 CD denotes 100
T5374 2525-2527 NN denotes mM
T5375 2528-2535 NN denotes glycine
T5376 2535-2537 , denotes ,
T5377 2537-2539 NN denotes pH
T5378 2540-2543 CD denotes 3.0
T5379 2543-2545 , denotes ,
T5380 2545-2548 CC denotes and
T5381 2549-2560 VBN denotes neutralized
T5382 2561-2572 RB denotes immediately
T5383 2572-2573 . denotes .
T5384 2573-2688 sentence denotes Using the affinity purified antibody, both human and mouse ADAM22 proteins were detected by Western blot analysis.
T5385 2574-2579 VBG denotes Using
T5387 2580-2583 DT denotes the
T5389 2584-2592 NN denotes affinity
T5390 2593-2601 VBN denotes purified
T5388 2602-2610 NN denotes antibody
T5391 2610-2612 , denotes ,
T5392 2612-2616 CC denotes both
T5394 2617-2622 JJ denotes human
T5395 2623-2626 CC denotes and
T5393 2627-2632 NN denotes mouse
T5397 2633-2639 NN denotes ADAM22
T5396 2640-2648 NN denotes proteins
T5398 2649-2653 VBD denotes were
T5386 2654-2662 VBN denotes detected
T5399 2663-2665 IN denotes by
T5400 2666-2673 NNP denotes Western
T5401 2674-2678 NN denotes blot
T5402 2679-2687 NN denotes analysis
T5403 2687-2688 . denotes .
T5404 2688-2792 sentence denotes However, unfortunately the antibody was not suitable for immunohistochemical analysis of mouse tissues.
T5405 2689-2696 RB denotes However
T5407 2696-2698 , denotes ,
T5408 2698-2711 RB denotes unfortunately
T5409 2712-2715 DT denotes the
T5410 2716-2724 NN denotes antibody
T5406 2725-2728 VBD denotes was
T5411 2729-2732 RB denotes not
T5412 2733-2741 JJ denotes suitable
T5413 2742-2745 IN denotes for
T5414 2746-2765 JJ denotes immunohistochemical
T5415 2766-2774 NN denotes analysis
T5416 2775-2777 IN denotes of
T5417 2778-2783 NN denotes mouse
T5418 2784-2791 NNS denotes tissues
T5419 2791-2792 . denotes .
T5499 2794-2801 NNP denotes Western
T5500 2802-2806 NN denotes blot
T5501 2807-2815 NN denotes analysis
T5502 2815-2908 sentence denotes The absence of ADAM22 protein in the Adam22 -/- mice was confirmed by Western blot analysis.
T5503 2816-2819 DT denotes The
T5504 2820-2827 NN denotes absence
T5506 2828-2830 IN denotes of
T5507 2831-2837 NN denotes ADAM22
T5508 2838-2845 NN denotes protein
T5509 2846-2848 IN denotes in
T5510 2849-2852 DT denotes the
T5512 2853-2859 NN denotes Adam22
T5513 2860-2861 SYM denotes -
T5514 2861-2862 HYPH denotes /
T5515 2862-2863 SYM denotes -
T5511 2864-2868 NNS denotes mice
T5516 2869-2872 VBD denotes was
T5505 2873-2882 VBN denotes confirmed
T5517 2883-2885 IN denotes by
T5518 2886-2893 NNP denotes Western
T5519 2894-2898 NN denotes blot
T5520 2899-2907 NN denotes analysis
T5521 2907-2908 . denotes .
T5522 2908-3145 sentence denotes Briefly, the cerebellum was isolated from a P14.5 mouse of each genotype and homogenized with a Polytron homogenizer in cell lysis buffer (50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 1% NP-40, Complete protease inhibitors [Roche Diagnostics]).
T5523 2909-2916 RB denotes Briefly
T5525 2916-2918 , denotes ,
T5526 2918-2921 DT denotes the
T5527 2922-2932 NN denotes cerebellum
T5528 2933-2936 VBD denotes was
T5524 2937-2945 VBN denotes isolated
T5529 2946-2950 IN denotes from
T5530 2951-2952 DT denotes a
T5532 2953-2958 NN denotes P14.5
T5531 2959-2964 NN denotes mouse
T5533 2965-2967 IN denotes of
T5534 2968-2972 DT denotes each
T5535 2973-2981 NN denotes genotype
T5536 2982-2985 CC denotes and
T5537 2986-2997 VBN denotes homogenized
T5538 2998-3002 IN denotes with
T5539 3003-3004 DT denotes a
T5541 3005-3013 NN denotes Polytron
T5540 3014-3025 NN denotes homogenizer
T5542 3026-3028 IN denotes in
T5543 3029-3033 NN denotes cell
T5544 3034-3039 NN denotes lysis
T5545 3040-3046 NN denotes buffer
T5546 3047-3048 -LRB- denotes (
T5548 3048-3050 CD denotes 50
T5549 3051-3053 NN denotes mM
T5551 3054-3058 NN denotes Tris
T5552 3058-3059 HYPH denotes -
T5550 3059-3062 NN denotes HCl
T5553 3062-3064 , denotes ,
T5554 3064-3066 NN denotes pH
T5555 3067-3070 CD denotes 7.5
T5556 3070-3072 , denotes ,
T5557 3072-3075 CD denotes 100
T5558 3076-3078 NN denotes mM
T5559 3079-3083 NN denotes NaCl
T5560 3083-3085 , denotes ,
T5561 3085-3086 CD denotes 1
T5562 3086-3087 NN denotes %
T5563 3088-3090 NN denotes NP
T5564 3090-3091 HYPH denotes -
T5565 3091-3093 CD denotes 40
T5566 3093-3095 , denotes ,
T5567 3095-3103 JJ denotes Complete
T5569 3104-3112 NN denotes protease
T5568 3113-3123 NNS denotes inhibitors
T5570 3124-3125 -LRB- denotes [
T5571 3125-3130 NNP denotes Roche
T5547 3131-3142 NNP denotes Diagnostics
T5572 3142-3143 -RRB- denotes ]
T5573 3143-3144 -RRB- denotes )
T5574 3144-3145 . denotes .
T5575 3145-3286 sentence denotes After removal of cell debris by centrifugation, the supernatant was separated on 10% SDS-PAGE, and transferred to a nitrocellulose membrane.
T5576 3146-3151 IN denotes After
T5578 3152-3159 NN denotes removal
T5579 3160-3162 IN denotes of
T5580 3163-3167 NN denotes cell
T5581 3168-3174 NN denotes debris
T5582 3175-3177 IN denotes by
T5583 3178-3192 NN denotes centrifugation
T5584 3192-3194 , denotes ,
T5585 3194-3197 DT denotes the
T5586 3198-3209 NN denotes supernatant
T5587 3210-3213 VBD denotes was
T5577 3214-3223 VBN denotes separated
T5588 3224-3226 IN denotes on
T5589 3227-3229 CD denotes 10
T5590 3229-3230 NN denotes %
T5592 3231-3234 NN denotes SDS
T5593 3234-3235 HYPH denotes -
T5591 3235-3239 NN denotes PAGE
T5594 3239-3241 , denotes ,
T5595 3241-3244 CC denotes and
T5596 3245-3256 VBN denotes transferred
T5597 3257-3259 IN denotes to
T5598 3260-3261 DT denotes a
T5600 3262-3276 NN denotes nitrocellulose
T5599 3277-3285 NN denotes membrane
T5601 3285-3286 . denotes .
T5602 3286-3394 sentence denotes The blot was then incubated with polyclonal antibody against the cytoplasmic domain of ADAM22 (at 1:1,000).
T5603 3287-3290 DT denotes The
T5604 3291-3295 NN denotes blot
T5606 3296-3299 VBD denotes was
T5607 3300-3304 RB denotes then
T5605 3305-3314 VBN denotes incubated
T5608 3315-3319 IN denotes with
T5609 3320-3330 JJ denotes polyclonal
T5610 3331-3339 NN denotes antibody
T5611 3340-3347 IN denotes against
T5612 3348-3351 DT denotes the
T5614 3352-3363 JJ denotes cytoplasmic
T5613 3364-3370 NN denotes domain
T5615 3371-3373 IN denotes of
T5616 3374-3380 NN denotes ADAM22
T5617 3381-3382 -LRB- denotes (
T5618 3382-3384 IN denotes at
T5619 3385-3386 CD denotes 1
T5620 3386-3387 SYM denotes :
T5621 3387-3392 CD denotes 1,000
T5622 3392-3393 -RRB- denotes )
T5623 3393-3394 . denotes .
T5624 3394-3560 sentence denotes Bound antibodies were visualized with horseradish peroxidase-labelled second antibody and a ECL-plus chemiluminescence detection system (Amersham Biosciences Corp.).
T5625 3395-3400 VBN denotes Bound
T5626 3401-3411 NNS denotes antibodies
T5628 3412-3416 VBD denotes were
T5627 3417-3427 VBN denotes visualized
T5629 3428-3432 IN denotes with
T5630 3433-3444 NN denotes horseradish
T5632 3445-3455 NN denotes peroxidase
T5634 3455-3456 HYPH denotes -
T5633 3456-3464 VBN denotes labelled
T5635 3465-3471 JJ denotes second
T5631 3472-3480 NN denotes antibody
T5636 3481-3484 CC denotes and
T5637 3485-3486 DT denotes a
T5639 3487-3490 NN denotes ECL
T5641 3490-3491 HYPH denotes -
T5640 3491-3495 NN denotes plus
T5642 3496-3513 NN denotes chemiluminescence
T5643 3514-3523 NN denotes detection
T5638 3524-3530 NN denotes system
T5644 3531-3532 -LRB- denotes (
T5646 3532-3540 NNP denotes Amersham
T5647 3541-3552 NNP denotes Biosciences
T5645 3553-3558 NNP denotes Corp.
T5648 3558-3559 -RRB- denotes )
T5649 3559-3560 . denotes .
T5703 3562-3569 JJ denotes Primary
T5705 3570-3577 NNP denotes Schwann
T5704 3578-3582 NN denotes cell
T5706 3583-3590 NN denotes culture
T5707 3590-3718 sentence denotes Primary Schwann cells were prepared from 4-month-old C57BL/6 mice according to Manent's protocol [32] with minor modifications.
T5708 3591-3598 JJ denotes Primary
T5710 3599-3606 NNP denotes Schwann
T5709 3607-3612 NNS denotes cells
T5712 3613-3617 VBD denotes were
T5711 3618-3626 VBN denotes prepared
T5713 3627-3631 IN denotes from
T5714 3632-3633 CD denotes 4
T5716 3633-3634 HYPH denotes -
T5715 3634-3639 NN denotes month
T5718 3639-3640 HYPH denotes -
T5717 3640-3643 JJ denotes old
T5720 3644-3649 NN denotes C57BL
T5721 3649-3650 HYPH denotes /
T5722 3650-3651 CD denotes 6
T5719 3652-3656 NNS denotes mice
T5723 3657-3666 VBG denotes according
T5724 3667-3669 IN denotes to
T5725 3670-3676 NNP denotes Manent
T5727 3676-3678 POS denotes 's
T5726 3679-3687 NN denotes protocol
T5728 3688-3689 -LRB- denotes [
T5729 3689-3691 CD denotes 32
T5730 3691-3692 -RRB- denotes ]
T5731 3693-3697 IN denotes with
T5732 3698-3703 JJ denotes minor
T5733 3704-3717 NNS denotes modifications
T5734 3717-3718 . denotes .
T5735 3718-4050 sentence denotes Briefly, sciatic nerves were removed and incubated for 7 days in the pre-treatment medium, which consisted of D-MEM (high glucose) supplemented with 10% FCS, 50 mg/ml gentamicin (Invitrogen Corp.), 2.5 μg/ml fungizone (Invitrogen Corp.), 2 μM forskolin (EMD Biosciences Inc.) and 10 ng/ml recombinant heregulin-beta1 (R&D Systems).
T5736 3719-3726 RB denotes Briefly
T5738 3726-3728 , denotes ,
T5739 3728-3735 JJ denotes sciatic
T5740 3736-3742 NNS denotes nerves
T5741 3743-3747 VBD denotes were
T5737 3748-3755 VBN denotes removed
T5742 3756-3759 CC denotes and
T5743 3760-3769 VBN denotes incubated
T5744 3770-3773 IN denotes for
T5745 3774-3775 CD denotes 7
T5746 3776-3780 NNS denotes days
T5747 3781-3783 IN denotes in
T5748 3784-3787 DT denotes the
T5750 3788-3801 JJ denotes pre-treatment
T5749 3802-3808 NN denotes medium
T5751 3808-3810 , denotes ,
T5752 3810-3815 WDT denotes which
T5753 3816-3825 VBD denotes consisted
T5754 3826-3828 IN denotes of
T5755 3829-3830 NN denotes D
T5757 3830-3831 HYPH denotes -
T5756 3831-3834 NN denotes MEM
T5758 3835-3836 -LRB- denotes (
T5760 3836-3840 JJ denotes high
T5759 3841-3848 NN denotes glucose
T5761 3848-3849 -RRB- denotes )
T5762 3850-3862 VBN denotes supplemented
T5763 3863-3867 IN denotes with
T5764 3868-3870 CD denotes 10
T5765 3870-3871 NN denotes %
T5766 3872-3875 NN denotes FCS
T5767 3875-3877 , denotes ,
T5768 3877-3879 CD denotes 50
T5769 3880-3882 NN denotes mg
T5771 3882-3883 SYM denotes /
T5772 3883-3885 NN denotes ml
T5770 3886-3896 NN denotes gentamicin
T5773 3897-3898 -LRB- denotes (
T5775 3898-3908 NNP denotes Invitrogen
T5774 3909-3914 NNP denotes Corp.
T5776 3914-3915 -RRB- denotes )
T5777 3915-3917 , denotes ,
T5778 3917-3920 CD denotes 2.5
T5779 3921-3923 NN denotes μg
T5781 3923-3924 SYM denotes /
T5782 3924-3926 NN denotes ml
T5780 3927-3936 NN denotes fungizone
T5783 3937-3938 -LRB- denotes (
T5785 3938-3948 NNP denotes Invitrogen
T5784 3949-3954 NNP denotes Corp.
T5786 3954-3955 -RRB- denotes )
T5787 3955-3957 , denotes ,
T5788 3957-3958 CD denotes 2
T5789 3959-3961 NN denotes μM
T5790 3962-3971 NN denotes forskolin
T5791 3972-3973 -LRB- denotes (
T5793 3973-3976 NNP denotes EMD
T5794 3977-3988 NNP denotes Biosciences
T5792 3989-3993 NNP denotes Inc.
T5795 3993-3994 -RRB- denotes )
T5796 3995-3998 CC denotes and
T5797 3999-4001 CD denotes 10
T5798 4002-4004 NN denotes ng
T5800 4004-4005 SYM denotes /
T5801 4005-4007 NN denotes ml
T5802 4008-4019 JJ denotes recombinant
T5803 4020-4029 NN denotes heregulin
T5804 4029-4030 HYPH denotes -
T5799 4030-4035 NN denotes beta1
T5805 4036-4037 -LRB- denotes (
T5807 4037-4038 NNP denotes R
T5808 4038-4039 CC denotes &
T5809 4039-4040 NNP denotes D
T5806 4041-4048 NNP denotes Systems
T5810 4048-4049 -RRB- denotes )
T5811 4049-4050 . denotes .
T5812 4050-4288 sentence denotes For dissociation, cultured sciatic nerves were cut into pieces and incubated at 37°C for 3 hours in Opti-MEM medium (Invitrogen Corp.) containing 130 U/ml collagenase type I (Invitrogen Corp.) and 0.4 U/ml dispase II (Roche Diagnostics).
T5813 4051-4054 IN denotes For
T5815 4055-4067 NN denotes dissociation
T5816 4067-4069 , denotes ,
T5817 4069-4077 VBN denotes cultured
T5819 4078-4085 JJ denotes sciatic
T5818 4086-4092 NNS denotes nerves
T5820 4093-4097 VBD denotes were
T5814 4098-4101 VBN denotes cut
T5821 4102-4106 IN denotes into
T5822 4107-4113 NNS denotes pieces
T5823 4114-4117 CC denotes and
T5824 4118-4127 VBN denotes incubated
T5825 4128-4130 IN denotes at
T5826 4131-4133 CD denotes 37
T5827 4133-4135 NN denotes °C
T5828 4136-4139 IN denotes for
T5829 4140-4141 CD denotes 3
T5830 4142-4147 NNS denotes hours
T5831 4148-4150 IN denotes in
T5832 4151-4155 NN denotes Opti
T5834 4155-4156 HYPH denotes -
T5833 4156-4159 NN denotes MEM
T5835 4160-4166 NN denotes medium
T5836 4167-4168 -LRB- denotes (
T5838 4168-4178 NNP denotes Invitrogen
T5837 4179-4184 NNP denotes Corp.
T5839 4184-4185 -RRB- denotes )
T5840 4186-4196 VBG denotes containing
T5841 4197-4200 CD denotes 130
T5842 4201-4202 NN denotes U
T5844 4202-4203 SYM denotes /
T5845 4203-4205 NN denotes ml
T5846 4206-4217 NN denotes collagenase
T5843 4218-4222 NN denotes type
T5847 4223-4224 CD denotes I
T5848 4225-4226 -LRB- denotes (
T5850 4226-4236 NNP denotes Invitrogen
T5849 4237-4242 NNP denotes Corp.
T5851 4242-4243 -RRB- denotes )
T5852 4244-4247 CC denotes and
T5853 4248-4251 CD denotes 0.4
T5854 4252-4253 NN denotes U
T5856 4253-4254 SYM denotes /
T5857 4254-4256 NN denotes ml
T5855 4257-4264 NN denotes dispase
T5858 4265-4267 CD denotes II
T5859 4268-4269 -LRB- denotes (
T5861 4269-4274 NNP denotes Roche
T5860 4275-4286 NNP denotes Diagnostics
T5862 4286-4287 -RRB- denotes )
T5863 4287-4288 . denotes .
T5864 4288-4418 sentence denotes Dissociated cells were resuspended in the pre-treatment medium and plated on Poly-D-Lysine/Laminin coated plate (BD Biosciences).
T5865 4289-4300 VBN denotes Dissociated
T5866 4301-4306 NNS denotes cells
T5868 4307-4311 VBD denotes were
T5867 4312-4323 VBN denotes resuspended
T5869 4324-4326 IN denotes in
T5870 4327-4330 DT denotes the
T5872 4331-4344 JJ denotes pre-treatment
T5871 4345-4351 NN denotes medium
T5873 4352-4355 CC denotes and
T5874 4356-4362 VBN denotes plated
T5875 4363-4365 IN denotes on
T5876 4366-4370 NN denotes Poly
T5878 4370-4371 HYPH denotes -
T5879 4371-4372 NN denotes D
T5880 4372-4373 HYPH denotes -
T5877 4373-4379 NN denotes Lysine
T5882 4379-4380 HYPH denotes /
T5883 4380-4387 NN denotes Laminin
T5881 4388-4394 VBN denotes coated
T5884 4395-4400 NN denotes plate
T5885 4401-4402 -LRB- denotes (
T5887 4402-4404 NN denotes BD
T5886 4405-4416 NNS denotes Biosciences
T5888 4416-4417 -RRB- denotes )
T5889 4417-4418 . denotes .
T5890 4418-4532 sentence denotes The purity of the cultured Schwann cells, as determined by indirect immunofluorescence analysis, approached 90 %.
T5891 4419-4422 DT denotes The
T5892 4423-4429 NN denotes purity
T5894 4430-4432 IN denotes of
T5895 4433-4436 DT denotes the
T5897 4437-4445 VBN denotes cultured
T5898 4446-4453 NNP denotes Schwann
T5896 4454-4459 NNS denotes cells
T5899 4459-4461 , denotes ,
T5900 4461-4463 IN denotes as
T5901 4464-4474 VBN denotes determined
T5902 4475-4477 IN denotes by
T5903 4478-4486 JJ denotes indirect
T5905 4487-4505 NN denotes immunofluorescence
T5904 4506-4514 NN denotes analysis
T5906 4514-4516 , denotes ,
T5893 4516-4526 VBD denotes approached
T5907 4527-4529 CD denotes 90
T5908 4530-4531 NN denotes %
T5909 4531-4532 . denotes .
T5980 4534-4536 NN denotes RT
T5982 4536-4537 HYPH denotes -
T5981 4537-4540 NN denotes PCR
T5983 4541-4549 NN denotes analysis
T5984 4549-4598 sentence denotes Adult C57BL/6 male mice were used in this study.
T5985 4550-4555 JJ denotes Adult
T5987 4556-4561 NN denotes C57BL
T5988 4561-4562 HYPH denotes /
T5989 4562-4563 CD denotes 6
T5990 4564-4568 JJ denotes male
T5986 4569-4573 NNS denotes mice
T5992 4574-4578 VBD denotes were
T5991 4579-4583 VBN denotes used
T5993 4584-4586 IN denotes in
T5994 4587-4591 DT denotes this
T5995 4592-4597 NN denotes study
T5996 4597-4598 . denotes .
T5997 4598-5001 sentence denotes Total RNAs purified from the cerebellum, spinal cord, sciatic nerve, DRG and cultured Schwann cells using TRIzol (Invitrogen Corp.) and RNeasy kit (Qiagen GmbH) were analysed by RT-PCR using SuperScript II and random primer (Invitrogen Corp.), and PCR amplification (40 cycles; 94°C-30 s, 60°C-30 s and 68°C-5 min) with Expand Hi-Fidelity DNA polymerase (Roche Diagnostics) and ADAM22-specific primers.
T5998 4599-4604 JJ denotes Total
T5999 4605-4609 NNS denotes RNAs
T6001 4610-4618 VBN denotes purified
T6002 4619-4623 IN denotes from
T6003 4624-4627 DT denotes the
T6004 4628-4638 NN denotes cerebellum
T6005 4638-4640 , denotes ,
T6006 4640-4646 JJ denotes spinal
T6007 4647-4651 NN denotes cord
T6008 4651-4653 , denotes ,
T6009 4653-4660 JJ denotes sciatic
T6010 4661-4666 NN denotes nerve
T6011 4666-4668 , denotes ,
T6012 4668-4671 NN denotes DRG
T6013 4672-4675 CC denotes and
T6014 4676-4684 VBN denotes cultured
T6016 4685-4692 NNP denotes Schwann
T6015 4693-4698 NNS denotes cells
T6017 4699-4704 VBG denotes using
T6018 4705-4711 NN denotes TRIzol
T6019 4712-4713 -LRB- denotes (
T6021 4713-4723 NNP denotes Invitrogen
T6020 4724-4729 NNP denotes Corp.
T6022 4729-4730 -RRB- denotes )
T6023 4731-4734 CC denotes and
T6024 4735-4741 NN denotes RNeasy
T6025 4742-4745 NN denotes kit
T6026 4746-4747 -LRB- denotes (
T6028 4747-4753 NNP denotes Qiagen
T6027 4754-4758 NNP denotes GmbH
T6029 4758-4759 -RRB- denotes )
T6030 4760-4764 VBD denotes were
T6000 4765-4773 VBN denotes analysed
T6031 4774-4776 IN denotes by
T6032 4777-4779 NN denotes RT
T6034 4779-4780 HYPH denotes -
T6033 4780-4783 NN denotes PCR
T6035 4784-4789 VBG denotes using
T6036 4790-4801 NNP denotes SuperScript
T6037 4802-4804 CD denotes II
T6038 4805-4808 CC denotes and
T6039 4809-4815 JJ denotes random
T6040 4816-4822 NN denotes primer
T6041 4823-4824 -LRB- denotes (
T6043 4824-4834 NNP denotes Invitrogen
T6042 4835-4840 NNP denotes Corp.
T6044 4840-4841 -RRB- denotes )
T6045 4841-4843 , denotes ,
T6046 4843-4846 CC denotes and
T6047 4847-4850 NN denotes PCR
T6048 4851-4864 NN denotes amplification
T6049 4865-4866 -LRB- denotes (
T6051 4866-4868 CD denotes 40
T6052 4869-4875 NNS denotes cycles
T6053 4875-4876 : denotes ;
T6054 4877-4879 CD denotes 94
T6050 4879-4881 NN denotes °C
T6055 4881-4882 HYPH denotes -
T6056 4882-4884 CD denotes 30
T6057 4885-4886 NN denotes s
T6058 4886-4888 , denotes ,
T6059 4888-4890 CD denotes 60
T6060 4890-4892 NN denotes °C
T6061 4892-4893 HYPH denotes -
T6062 4893-4895 CD denotes 30
T6063 4896-4897 NN denotes s
T6064 4898-4901 CC denotes and
T6065 4902-4904 CD denotes 68
T6066 4904-4906 NN denotes °C
T6067 4906-4907 HYPH denotes -
T6068 4907-4908 CD denotes 5
T6069 4909-4912 NN denotes min
T6070 4912-4913 -RRB- denotes )
T6071 4914-4918 IN denotes with
T6072 4919-4925 NNP denotes Expand
T6074 4926-4928 NNP denotes Hi
T6076 4928-4929 HYPH denotes -
T6075 4929-4937 NNP denotes Fidelity
T6077 4938-4941 NN denotes DNA
T6073 4942-4952 NN denotes polymerase
T6078 4953-4954 -LRB- denotes (
T6080 4954-4959 NNP denotes Roche
T6079 4960-4971 NNP denotes Diagnostics
T6081 4971-4972 -RRB- denotes )
T6082 4973-4976 CC denotes and
T6083 4977-4983 NN denotes ADAM22
T6085 4983-4984 HYPH denotes -
T6084 4984-4992 JJ denotes specific
T6086 4993-5000 NNS denotes primers
T6087 5000-5001 . denotes .
T6088 5001-5180 sentence denotes To detect splicing variants in the cytoplasmic domain, a forward primer was placed just upstream of the transmembrane domain and a reverse primer was set on the terminating exon.
T6089 5002-5004 TO denotes To
T6090 5005-5011 VB denotes detect
T6092 5012-5020 NN denotes splicing
T6093 5021-5029 NNS denotes variants
T6094 5030-5032 IN denotes in
T6095 5033-5036 DT denotes the
T6097 5037-5048 JJ denotes cytoplasmic
T6096 5049-5055 NN denotes domain
T6098 5055-5057 , denotes ,
T6099 5057-5058 DT denotes a
T6101 5059-5066 JJ denotes forward
T6100 5067-5073 NN denotes primer
T6102 5074-5077 VBD denotes was
T6091 5078-5084 VBN denotes placed
T6103 5085-5089 RB denotes just
T6104 5090-5098 RB denotes upstream
T6105 5099-5101 IN denotes of
T6106 5102-5105 DT denotes the
T6108 5106-5119 JJ denotes transmembrane
T6107 5120-5126 NN denotes domain
T6109 5127-5130 CC denotes and
T6110 5131-5132 DT denotes a
T6112 5133-5140 JJ denotes reverse
T6111 5141-5147 NN denotes primer
T6114 5148-5151 VBD denotes was
T6113 5152-5155 VBN denotes set
T6115 5156-5158 IN denotes on
T6116 5159-5162 DT denotes the
T6118 5163-5174 VBG denotes terminating
T6117 5175-5179 NN denotes exon
T6119 5179-5180 . denotes .
T6120 5180-5228 sentence denotes The primers used in this study were as follows.
T6121 5181-5184 DT denotes The
T6122 5185-5192 NNS denotes primers
T6124 5193-5197 VBN denotes used
T6125 5198-5200 IN denotes in
T6126 5201-5205 DT denotes this
T6127 5206-5211 NN denotes study
T6123 5212-5216 VBD denotes were
T6128 5217-5219 IN denotes as
T6129 5220-5227 VBZ denotes follows
T6130 5227-5228 . denotes .
T6131 5228-5325 sentence denotes ISH04-forward: 5'-AACAGGCACTGGACAGGGGCTGAC-3' and ISH04-reverse: 5'-AATGGATGTCTCCCATAGCCTGGC-3'.
T6132 5229-5234 NN denotes ISH04
T6133 5234-5235 HYPH denotes -
T6134 5235-5242 JJ denotes forward
T6135 5242-5244 : denotes :
T6136 5244-5245 CD denotes 5
T6138 5245-5246 SYM denotes '
T6139 5246-5247 HYPH denotes -
T6137 5247-5271 NN denotes AACAGGCACTGGACAGGGGCTGAC
T6140 5271-5272 HYPH denotes -
T6141 5272-5273 CD denotes 3
T6142 5273-5274 SYM denotes '
T6143 5275-5278 CC denotes and
T6144 5279-5284 NN denotes ISH04
T6145 5284-5285 HYPH denotes -
T6146 5285-5292 JJ denotes reverse
T6147 5292-5294 : denotes :
T6148 5294-5295 CD denotes 5
T6150 5295-5296 SYM denotes '
T6151 5296-5297 HYPH denotes -
T6149 5297-5321 NN denotes AATGGATGTCTCCCATAGCCTGGC
T6152 5321-5322 HYPH denotes -
T6153 5322-5323 CD denotes 3
T6154 5323-5324 SYM denotes '
T6155 5324-5325 . denotes .
T6212 5327-5341 NN denotes Histopathology
T6213 5341-5382 sentence denotes Mice were anesthetized with ethyl ether.
T6214 5342-5346 NNS denotes Mice
T6216 5347-5351 VBD denotes were
T6215 5352-5364 VBN denotes anesthetized
T6217 5365-5369 IN denotes with
T6218 5370-5375 NN denotes ethyl
T6219 5376-5381 NN denotes ether
T6220 5381-5382 . denotes .
T6221 5382-5501 sentence denotes Whole-body perfusion by 2% paraformaldehyde-glutaraldehyde solution followed by heparin-included saline was performed.
T6222 5383-5388 JJ denotes Whole
T6224 5388-5389 HYPH denotes -
T6223 5389-5393 NN denotes body
T6225 5394-5403 NN denotes perfusion
T6227 5404-5406 IN denotes by
T6228 5407-5408 CD denotes 2
T6229 5408-5409 NN denotes %
T6231 5410-5426 NN denotes paraformaldehyde
T6233 5426-5427 HYPH denotes -
T6232 5427-5441 NN denotes glutaraldehyde
T6230 5442-5450 NN denotes solution
T6234 5451-5459 VBN denotes followed
T6235 5460-5462 IN denotes by
T6236 5463-5470 NN denotes heparin
T6238 5470-5471 HYPH denotes -
T6237 5471-5479 VBN denotes included
T6239 5480-5486 NN denotes saline
T6240 5487-5490 VBD denotes was
T6226 5491-5500 VBN denotes performed
T6241 5500-5501 . denotes .
T6242 5501-5615 sentence denotes Sciatic nerves, trigeminal nerves, brain and spinal cord were removed and fixed in 10% neutral-buffered formalin.
T6243 5502-5509 JJ denotes Sciatic
T6244 5510-5516 NNS denotes nerves
T6246 5516-5518 , denotes ,
T6247 5518-5528 JJ denotes trigeminal
T6248 5529-5535 NNS denotes nerves
T6249 5535-5537 , denotes ,
T6250 5537-5542 NN denotes brain
T6251 5543-5546 CC denotes and
T6252 5547-5553 JJ denotes spinal
T6253 5554-5558 NN denotes cord
T6254 5559-5563 VBD denotes were
T6245 5564-5571 VBN denotes removed
T6255 5572-5575 CC denotes and
T6256 5576-5581 VBN denotes fixed
T6257 5582-5584 IN denotes in
T6258 5585-5587 CD denotes 10
T6259 5587-5588 NN denotes %
T6261 5589-5596 JJ denotes neutral
T6263 5596-5597 HYPH denotes -
T6262 5597-5605 VBN denotes buffered
T6260 5606-5614 NN denotes formalin
T6264 5614-5615 . denotes .
T6265 5615-5763 sentence denotes The spinal cord and other nerve blocks were washed and postfixed with 2% osmium tetroxide, and were dehydrated in ethanol and equilibrated in Epon.
T6266 5616-5619 DT denotes The
T6268 5620-5626 JJ denotes spinal
T6267 5627-5631 NN denotes cord
T6270 5632-5635 CC denotes and
T6271 5636-5641 JJ denotes other
T6273 5642-5647 NN denotes nerve
T6272 5648-5654 NNS denotes blocks
T6274 5655-5659 VBD denotes were
T6269 5660-5666 VBN denotes washed
T6275 5667-5670 CC denotes and
T6276 5671-5680 VBN denotes postfixed
T6277 5681-5685 IN denotes with
T6278 5686-5687 CD denotes 2
T6279 5687-5688 NN denotes %
T6281 5689-5695 NN denotes osmium
T6280 5696-5705 NN denotes tetroxide
T6282 5705-5707 , denotes ,
T6283 5707-5710 CC denotes and
T6284 5711-5715 VBD denotes were
T6285 5716-5726 VBN denotes dehydrated
T6286 5727-5729 IN denotes in
T6287 5730-5737 NN denotes ethanol
T6288 5738-5741 CC denotes and
T6289 5742-5754 VBN denotes equilibrated
T6290 5755-5757 IN denotes in
T6291 5758-5762 NN denotes Epon
T6292 5762-5763 . denotes .
T6293 5763-5881 sentence denotes Epon embedded semithin sections were stained with toluidine blue and were subjected to light microscopic examination.
T6294 5764-5768 NN denotes Epon
T6295 5769-5777 VBN denotes embedded
T6297 5778-5786 JJ denotes semithin
T6296 5787-5795 NNS denotes sections
T6299 5796-5800 VBD denotes were
T6298 5801-5808 VBN denotes stained
T6300 5809-5813 IN denotes with
T6301 5814-5823 NN denotes toluidine
T6302 5824-5828 NN denotes blue
T6303 5829-5832 CC denotes and
T6304 5833-5837 VBD denotes were
T6305 5838-5847 VBN denotes subjected
T6306 5848-5850 IN denotes to
T6307 5851-5856 NN denotes light
T6308 5857-5868 JJ denotes microscopic
T6309 5869-5880 NN denotes examination
T6310 5880-5881 . denotes .
T6311 5881-6074 sentence denotes For electron microscopic analysis, thin sections were cut using an ultramicrotome, stained with 1.5% uranylacetate in 50% ethanol and 0.8% lead citrate, and analysed using electron microscope.
T6312 5882-5885 IN denotes For
T6314 5886-5894 NN denotes electron
T6315 5895-5906 JJ denotes microscopic
T6316 5907-5915 NN denotes analysis
T6317 5915-5917 , denotes ,
T6318 5917-5921 JJ denotes thin
T6319 5922-5930 NNS denotes sections
T6320 5931-5935 VBD denotes were
T6313 5936-5939 VBN denotes cut
T6321 5940-5945 VBG denotes using
T6322 5946-5948 DT denotes an
T6323 5949-5963 NN denotes ultramicrotome
T6324 5963-5965 , denotes ,
T6325 5965-5972 VBN denotes stained
T6326 5973-5977 IN denotes with
T6327 5978-5981 CD denotes 1.5
T6328 5981-5982 NN denotes %
T6329 5983-5996 NN denotes uranylacetate
T6330 5997-5999 IN denotes in
T6331 6000-6002 CD denotes 50
T6332 6002-6003 NN denotes %
T6333 6004-6011 NN denotes ethanol
T6334 6012-6015 CC denotes and
T6335 6016-6019 CD denotes 0.8
T6336 6019-6020 NN denotes %
T6338 6021-6025 NN denotes lead
T6337 6026-6033 NN denotes citrate
T6339 6033-6035 , denotes ,
T6340 6035-6038 CC denotes and
T6341 6039-6047 VBN denotes analysed
T6342 6048-6053 VBG denotes using
T6343 6054-6062 NN denotes electron
T6344 6063-6073 NN denotes microscope
T6345 6073-6074 . denotes .
T6346 6074-6163 sentence denotes All procedures were conducted according to the Eisai Animal Care Committee's guidelines.
T6347 6075-6078 DT denotes All
T6348 6079-6089 NNS denotes procedures
T6350 6090-6094 VBD denotes were
T6349 6095-6104 VBN denotes conducted
T6351 6105-6114 VBG denotes according
T6352 6115-6117 IN denotes to
T6353 6118-6121 DT denotes the
T6355 6122-6127 NNP denotes Eisai
T6356 6128-6134 NNP denotes Animal
T6357 6135-6139 NNP denotes Care
T6354 6140-6149 NNP denotes Committee
T6359 6149-6151 POS denotes 's
T6358 6152-6162 NNS denotes guidelines
T6360 6162-6163 . denotes .
T6425 6165-6185 NN denotes Immunohistochemistry
T6426 6185-6322 sentence denotes Frozen sections were rinsed in 0.1% Triton X-100/PBS at room temperature for 1 hour, and were blocked in BLOCKACE solution (Dai-nippon).
T6427 6186-6192 JJ denotes Frozen
T6428 6193-6201 NNS denotes sections
T6430 6202-6206 VBD denotes were
T6429 6207-6213 VBN denotes rinsed
T6431 6214-6216 IN denotes in
T6432 6217-6220 CD denotes 0.1
T6433 6220-6221 NN denotes %
T6434 6222-6228 NN denotes Triton
T6435 6229-6230 NN denotes X
T6436 6230-6231 HYPH denotes -
T6437 6231-6234 CD denotes 100
T6438 6234-6235 HYPH denotes /
T6439 6235-6238 NN denotes PBS
T6440 6239-6241 IN denotes at
T6441 6242-6246 NN denotes room
T6442 6247-6258 NN denotes temperature
T6443 6259-6262 IN denotes for
T6444 6263-6264 CD denotes 1
T6445 6265-6269 NN denotes hour
T6446 6269-6271 , denotes ,
T6447 6271-6274 CC denotes and
T6448 6275-6279 VBD denotes were
T6449 6280-6287 VBN denotes blocked
T6450 6288-6290 IN denotes in
T6451 6291-6299 NN denotes BLOCKACE
T6452 6300-6308 NN denotes solution
T6453 6309-6310 -LRB- denotes (
T6455 6310-6313 NNP denotes Dai
T6456 6313-6314 HYPH denotes -
T6454 6314-6320 NNP denotes nippon
T6457 6320-6321 -RRB- denotes )
T6458 6321-6322 . denotes .
T6459 6322-6397 sentence denotes The following antibodies were incubated overnight in 0.1% BLOCKACE at 4°C:
T6460 6323-6326 DT denotes The
T6462 6327-6336 VBG denotes following
T6461 6337-6347 NNS denotes antibodies
T6464 6348-6352 VBD denotes were
T6463 6353-6362 VBN denotes incubated
T6465 6363-6372 RB denotes overnight
T6466 6373-6375 IN denotes in
T6467 6376-6379 CD denotes 0.1
T6468 6379-6380 NN denotes %
T6469 6381-6389 NN denotes BLOCKACE
T6470 6390-6392 IN denotes at
T6471 6393-6394 CD denotes 4
T6472 6394-6396 NN denotes °C
T6473 6396-6397 : denotes :
T6474 6397-6612 sentence denotes monoclonal mouse anti-calbindin 28 K (Sigma, 1:200); monoclonal mouse anti-MBP (SMI-99, Sternberger Monoclonals Inc., 1:50); monoclonal mouse anti-Neu N (CHEMICON, 1:100); rabbit anti-S100 polyclonal (DAKO, 1:200).
T6475 6398-6408 JJ denotes monoclonal
T6477 6409-6414 NN denotes mouse
T6478 6415-6429 JJ denotes anti-calbindin
T6479 6430-6432 CD denotes 28
T6476 6433-6434 NN denotes K
T6480 6435-6436 -LRB- denotes (
T6482 6436-6441 NNP denotes Sigma
T6483 6441-6443 , denotes ,
T6484 6443-6444 CD denotes 1
T6485 6444-6445 : denotes :
T6481 6445-6448 CD denotes 200
T6486 6448-6449 -RRB- denotes )
T6487 6449-6450 : denotes ;
T6488 6451-6461 JJ denotes monoclonal
T6489 6462-6467 NN denotes mouse
T6490 6468-6476 JJ denotes anti-MBP
T6491 6477-6478 -LRB- denotes (
T6493 6478-6481 NNP denotes SMI
T6494 6481-6482 HYPH denotes -
T6495 6482-6484 CD denotes 99
T6496 6484-6486 , denotes ,
T6497 6486-6497 NNP denotes Sternberger
T6498 6498-6509 NNP denotes Monoclonals
T6499 6510-6514 NNP denotes Inc.
T6500 6514-6516 , denotes ,
T6501 6516-6517 CD denotes 1
T6502 6517-6518 SYM denotes :
T6492 6518-6520 CD denotes 50
T6503 6520-6521 -RRB- denotes )
T6504 6521-6522 : denotes ;
T6505 6523-6533 JJ denotes monoclonal
T6507 6534-6539 NN denotes mouse
T6508 6540-6548 JJ denotes anti-Neu
T6506 6549-6550 NN denotes N
T6509 6551-6552 -LRB- denotes (
T6511 6552-6560 NNP denotes CHEMICON
T6512 6560-6562 , denotes ,
T6513 6562-6563 CD denotes 1
T6514 6563-6564 SYM denotes :
T6510 6564-6567 CD denotes 100
T6515 6567-6568 -RRB- denotes )
T6516 6568-6569 : denotes ;
T6517 6570-6576 NN denotes rabbit
T6518 6577-6586 JJ denotes anti-S100
T6519 6587-6597 JJ denotes polyclonal
T6520 6598-6599 -LRB- denotes (
T6522 6599-6603 NNP denotes DAKO
T6523 6603-6605 , denotes ,
T6524 6605-6606 CD denotes 1
T6525 6606-6607 SYM denotes :
T6521 6607-6610 CD denotes 200
T6526 6610-6611 -RRB- denotes )
T6527 6611-6612 . denotes .
T6528 6612-6666 sentence denotes Nuclei were counterstained with 1 μg/ml DAPI (Sigma).
T6529 6613-6619 NNS denotes Nuclei
T6531 6620-6624 VBD denotes were
T6530 6625-6639 VBN denotes counterstained
T6532 6640-6644 IN denotes with
T6533 6645-6646 CD denotes 1
T6534 6647-6649 NN denotes μg
T6536 6649-6650 SYM denotes /
T6537 6650-6652 NN denotes ml
T6535 6653-6657 NN denotes DAPI
T6538 6658-6659 -LRB- denotes (
T6539 6659-6664 NNP denotes Sigma
T6540 6664-6665 -RRB- denotes )
T6541 6665-6666 . denotes .
T6542 6666-6763 sentence denotes MBP staining was performed after microwave epitope retrieval in 10 mmol/L citrate buffer (pH 6).
T6543 6667-6670 NN denotes MBP
T6544 6671-6679 NN denotes staining
T6546 6680-6683 VBD denotes was
T6545 6684-6693 VBN denotes performed
T6547 6694-6699 IN denotes after
T6548 6700-6709 NN denotes microwave
T6550 6710-6717 NN denotes epitope
T6549 6718-6727 NN denotes retrieval
T6551 6728-6730 IN denotes in
T6552 6731-6733 CD denotes 10
T6553 6734-6738 NN denotes mmol
T6555 6738-6739 SYM denotes /
T6556 6739-6740 NN denotes L
T6557 6741-6748 NN denotes citrate
T6554 6749-6755 NN denotes buffer
T6558 6756-6757 -LRB- denotes (
T6559 6757-6759 NN denotes pH
T6560 6760-6761 CD denotes 6
T6561 6761-6762 -RRB- denotes )
T6562 6762-6763 . denotes .
T6563 6763-6964 sentence denotes Sections were incubated for 1 hour with secondary antibodies: Cy3-labeled donkey anti-rabbit IgG (Jackson Immuno Research, 1:200), FITC-labelled donkey anti-mouse IgG (Jackson Immuno Research, 1:200).
T6564 6764-6772 NNS denotes Sections
T6566 6773-6777 VBD denotes were
T6565 6778-6787 VBN denotes incubated
T6567 6788-6791 IN denotes for
T6568 6792-6793 CD denotes 1
T6569 6794-6798 NN denotes hour
T6570 6799-6803 IN denotes with
T6571 6804-6813 JJ denotes secondary
T6572 6814-6824 NNS denotes antibodies
T6573 6824-6826 : denotes :
T6574 6826-6829 NN denotes Cy3
T6576 6829-6830 HYPH denotes -
T6575 6830-6837 VBN denotes labeled
T6578 6838-6844 NN denotes donkey
T6579 6845-6856 JJ denotes anti-rabbit
T6577 6857-6860 NN denotes IgG
T6580 6861-6862 -LRB- denotes (
T6582 6862-6869 NNP denotes Jackson
T6584 6870-6876 NNP denotes Immuno
T6583 6877-6885 NNP denotes Research
T6585 6885-6887 , denotes ,
T6586 6887-6888 CD denotes 1
T6587 6888-6889 SYM denotes :
T6581 6889-6892 CD denotes 200
T6588 6892-6893 -RRB- denotes )
T6589 6893-6895 , denotes ,
T6590 6895-6899 NN denotes FITC
T6592 6899-6900 HYPH denotes -
T6591 6900-6908 VBN denotes labelled
T6594 6909-6915 NN denotes donkey
T6595 6916-6926 JJ denotes anti-mouse
T6593 6927-6930 NN denotes IgG
T6596 6931-6932 -LRB- denotes (
T6598 6932-6939 NNP denotes Jackson
T6600 6940-6946 NNP denotes Immuno
T6599 6947-6955 NNP denotes Research
T6601 6955-6957 , denotes ,
T6602 6957-6958 CD denotes 1
T6603 6958-6959 SYM denotes :
T6597 6959-6962 CD denotes 200
T6604 6962-6963 -RRB- denotes )
T6605 6963-6964 . denotes .
T6606 6964-7077 sentence denotes Sections were photographed with an Olympus microscope (Olympus IX71) equipped with a high-resolution CCD camera.
T6607 6965-6973 NNS denotes Sections
T6609 6974-6978 VBD denotes were
T6608 6979-6991 VBN denotes photographed
T6610 6992-6996 IN denotes with
T6611 6997-6999 DT denotes an
T6613 7000-7007 NNP denotes Olympus
T6612 7008-7018 NN denotes microscope
T6614 7019-7020 -LRB- denotes (
T6616 7020-7027 NNP denotes Olympus
T6615 7028-7032 NN denotes IX71
T6617 7032-7033 -RRB- denotes )
T6618 7034-7042 VBN denotes equipped
T6619 7043-7047 IN denotes with
T6620 7048-7049 DT denotes a
T6622 7050-7054 JJ denotes high
T6624 7054-7055 HYPH denotes -
T6623 7055-7065 NN denotes resolution
T6625 7066-7069 NN denotes CCD
T6621 7070-7076 NN denotes camera
T6626 7076-7077 . denotes .
T6692 7079-7082 NN denotes RNA
T6694 7083-7085 FW denotes in
T6695 7086-7090 FW denotes situ
T6693 7091-7104 NN denotes hybridisation
T6696 7104-7193 sentence denotes In situ hybridisation on frozen sections were carried out using 35S-labelled RNA probes.
T6697 7105-7107 FW denotes In
T6698 7108-7112 FW denotes situ
T6699 7113-7126 NN denotes hybridisation
T6701 7127-7129 IN denotes on
T6702 7130-7136 VBN denotes frozen
T6703 7137-7145 NNS denotes sections
T6704 7146-7150 VBD denotes were
T6700 7151-7158 VBN denotes carried
T6705 7159-7162 RP denotes out
T6706 7163-7168 VBG denotes using
T6707 7169-7172 CD denotes 35S
T6709 7172-7173 HYPH denotes -
T6708 7173-7181 VBN denotes labelled
T6711 7182-7185 NN denotes RNA
T6710 7186-7192 NNS denotes probes
T6712 7192-7193 . denotes .
T6713 7193-7349 sentence denotes Briefly, 610 bp of mouse ADAM22 cDNA (position: 1351–1960 from initiating ATG) was cloned into the pBluescript II SK(+) or the pBluescript II KS(+) vector.
T6714 7194-7201 RB denotes Briefly
T6716 7201-7203 , denotes ,
T6717 7203-7206 CD denotes 610
T6718 7207-7209 NN denotes bp
T6719 7210-7212 IN denotes of
T6720 7213-7218 NN denotes mouse
T6722 7219-7225 NN denotes ADAM22
T6721 7226-7230 NN denotes cDNA
T6723 7231-7232 -LRB- denotes (
T6725 7232-7240 NN denotes position
T6726 7240-7242 : denotes :
T6727 7242-7246 CD denotes 1351
T6728 7246-7247 SYM denotes –
T6724 7247-7251 CD denotes 1960
T6729 7252-7256 IN denotes from
T6730 7257-7267 VBG denotes initiating
T6731 7268-7271 NN denotes ATG
T6732 7271-7272 -RRB- denotes )
T6733 7273-7276 VBD denotes was
T6715 7277-7283 VBN denotes cloned
T6734 7284-7288 IN denotes into
T6735 7289-7292 DT denotes the
T6737 7293-7304 NNP denotes pBluescript
T6738 7305-7307 CD denotes II
T6736 7308-7310 NN denotes SK
T6739 7310-7311 -LRB- denotes (
T6740 7311-7312 SYM denotes +
T6741 7312-7313 -RRB- denotes )
T6742 7314-7316 CC denotes or
T6743 7317-7320 DT denotes the
T6745 7321-7332 NNP denotes pBluescript
T6746 7333-7335 CD denotes II
T6744 7336-7338 NN denotes KS
T6748 7338-7339 -LRB- denotes (
T6749 7339-7340 SYM denotes +
T6750 7340-7341 -RRB- denotes )
T6747 7342-7348 NN denotes vector
T6751 7348-7349 . denotes .
T6752 7349-7475 sentence denotes Using these plasmids as templates, sense and antisense labelled RNA probes were generated by T7 RNA polymerase and [α35S]UTP.
T6753 7350-7355 VBG denotes Using
T6755 7356-7361 DT denotes these
T6756 7362-7370 NNS denotes plasmids
T6757 7371-7373 IN denotes as
T6758 7374-7383 NNS denotes templates
T6759 7383-7385 , denotes ,
T6760 7385-7390 NN denotes sense
T6762 7391-7394 CC denotes and
T6763 7395-7404 JJ denotes antisense
T6761 7405-7413 VBN denotes labelled
T6765 7414-7417 NN denotes RNA
T6764 7418-7424 NNS denotes probes
T6766 7425-7429 VBD denotes were
T6754 7430-7439 VBN denotes generated
T6767 7440-7442 IN denotes by
T6768 7443-7445 NN denotes T7
T6770 7446-7449 NN denotes RNA
T6769 7450-7460 NN denotes polymerase
T6771 7461-7464 CC denotes and
T6772 7465-7466 -LRB- denotes [
T6774 7466-7470 NN denotes α35S
T6775 7470-7471 -RRB- denotes ]
T6773 7471-7474 NN denotes UTP
T6776 7474-7475 . denotes .
T6777 7475-7569 sentence denotes Frozen brain and spinal cord from 2 month-old C57BL/6 female mice were used in this analysis.
T6778 7476-7482 VBN denotes Frozen
T6779 7483-7488 NN denotes brain
T6781 7489-7492 CC denotes and
T6782 7493-7499 JJ denotes spinal
T6783 7500-7504 NN denotes cord
T6784 7505-7509 IN denotes from
T6785 7510-7511 CD denotes 2
T6786 7512-7517 NN denotes month
T6788 7517-7518 HYPH denotes -
T6787 7518-7521 JJ denotes old
T6790 7522-7527 NN denotes C57BL
T6791 7527-7528 HYPH denotes /
T6792 7528-7529 CD denotes 6
T6793 7530-7536 JJ denotes female
T6789 7537-7541 NNS denotes mice
T6794 7542-7546 VBD denotes were
T6780 7547-7551 VBN denotes used
T6795 7552-7554 IN denotes in
T6796 7555-7559 DT denotes this
T6797 7560-7568 NN denotes analysis
T6798 7568-7569 . denotes .
T6799 7569-7699 sentence denotes Pretreatment, hybridisation, RNase treatment and washing was carried out following the protocol described in the literature [33].
T6800 7570-7582 NN denotes Pretreatment
T6802 7582-7584 , denotes ,
T6803 7584-7597 NN denotes hybridisation
T6804 7597-7599 , denotes ,
T6805 7599-7604 NN denotes RNase
T6806 7605-7614 NN denotes treatment
T6807 7615-7618 CC denotes and
T6808 7619-7626 NN denotes washing
T6809 7627-7630 VBD denotes was
T6801 7631-7638 VBN denotes carried
T6810 7639-7642 RP denotes out
T6811 7643-7652 VBG denotes following
T6812 7653-7656 DT denotes the
T6813 7657-7665 NN denotes protocol
T6814 7666-7675 VBN denotes described
T6815 7676-7678 IN denotes in
T6816 7679-7682 DT denotes the
T6817 7683-7693 NN denotes literature
T6818 7694-7695 -LRB- denotes [
T6819 7695-7697 CD denotes 33
T6820 7697-7698 -RRB- denotes ]
T6821 7698-7699 . denotes .
T6822 7699-7854 sentence denotes Dehydrated slides were attached to imaging plates for 48 hours and the autoradiograms were analysed using a Bio-Image Analyser (BAS3000, Fuji Photo Film).
T6823 7700-7710 VBN denotes Dehydrated
T6824 7711-7717 NNS denotes slides
T6826 7718-7722 VBD denotes were
T6825 7723-7731 VBN denotes attached
T6827 7732-7734 IN denotes to
T6828 7735-7742 NN denotes imaging
T6829 7743-7749 NNS denotes plates
T6830 7750-7753 IN denotes for
T6831 7754-7756 CD denotes 48
T6832 7757-7762 NNS denotes hours
T6833 7763-7766 CC denotes and
T6834 7767-7770 DT denotes the
T6835 7771-7785 NNS denotes autoradiograms
T6837 7786-7790 VBD denotes were
T6836 7791-7799 VBN denotes analysed
T6838 7800-7805 VBG denotes using
T6839 7806-7807 DT denotes a
T6841 7808-7817 NNP denotes Bio-Image
T6840 7818-7826 NNP denotes Analyser
T6842 7827-7828 -LRB- denotes (
T6844 7828-7835 NN denotes BAS3000
T6845 7835-7837 , denotes ,
T6846 7837-7841 NNP denotes Fuji
T6847 7842-7847 NNP denotes Photo
T6843 7848-7852 NNP denotes Film
T6848 7852-7853 -RRB- denotes )
T6849 7853-7854 . denotes .
R2896 T4652 T4651 prep of,Construction
R2897 T4653 T4654 det an,vector
R2898 T4654 T4652 pobj vector,of
R2899 T4655 T4654 nmod Adam22,vector
R2900 T4656 T4657 npadvmod gene,targeting
R2901 T4657 T4654 amod targeting,vector
R2902 T4658 T4657 punct -,targeting
R2903 T4660 T4661 det The,fragments
R2904 T4661 T4667 nsubjpass fragments,amplified
R2905 T4662 T4663 nummod 18,kb
R2906 T4663 T4661 nmod kb,fragments
R2907 T4664 T4663 punct -,kb
R2908 T4665 T4661 amod genomic,fragments
R2909 T4666 T4661 compound DNA,fragments
R2910 T4668 T4661 acl covering,fragments
R2911 T4669 T4670 nmod exons,5
R2912 T4670 T4668 dobj 5,covering
R2913 T4671 T4670 punct ", ",5
R2914 T4672 T4670 conj 6,5
R2915 T4673 T4672 punct ", ",6
R2916 T4674 T4672 conj 7,6
R2917 T4675 T4674 punct ", ",7
R2918 T4676 T4674 conj 8,7
R2919 T4677 T4676 cc and,8
R2920 T4678 T4676 conj 9,8
R2921 T4679 T4670 prep of,5
R2922 T4680 T4681 det the,gene
R2923 T4681 T4679 pobj gene,of
R2924 T4682 T4681 compound Adam22,gene
R2925 T4683 T4667 auxpass were,amplified
R2926 T4684 T4667 prep from,amplified
R2927 T4685 T4686 nmod C57BL,DNA
R2928 T4686 T4684 pobj DNA,from
R2929 T4687 T4685 punct /,C57BL
R2930 T4688 T4685 nummod 6,C57BL
R2931 T4689 T4686 amod genomic,DNA
R2932 T4690 T4667 prep by,amplified
R2933 T4691 T4690 pobj PCR,by
R2934 T4692 T4667 advcl using,amplified
R2935 T4693 T4694 compound Expand,mix
R2936 T4694 T4692 dobj mix,using
R2937 T4695 T4696 compound Hi,Fidelity
R2938 T4696 T4694 compound Fidelity,mix
R2939 T4697 T4696 punct -,Fidelity
R2940 T4698 T4694 compound enzyme,mix
R2941 T4699 T4700 punct (,Diagnostics
R2942 T4700 T4694 parataxis Diagnostics,mix
R2943 T4701 T4700 compound Roche,Diagnostics
R2944 T4702 T4700 punct ),Diagnostics
R2945 T4703 T4694 cc and,mix
R2946 T4704 T4694 conj primers,mix
R2947 T4705 T4704 acl designed,primers
R2948 T4706 T4705 prep for,designed
R2949 T4707 T4708 det each,exon
R2950 T4708 T4706 pobj exon,for
R2951 T4709 T4667 punct .,amplified
R2952 T4711 T4712 aux To,generate
R2953 T4712 T4713 advcl generate,inserted
R2954 T4714 T4715 det a,mutation
R2955 T4715 T4712 dobj mutation,generate
R2956 T4716 T4712 prep in,generate
R2957 T4717 T4718 det the,gene
R2958 T4718 T4716 pobj gene,in
R2959 T4719 T4718 compound mouse,gene
R2960 T4720 T4718 compound Adam22,gene
R2961 T4721 T4713 punct ", ",inserted
R2962 T4722 T4713 nsubj we,inserted
R2963 T4723 T4724 det the,cassette
R2964 T4724 T4713 dobj cassette,inserted
R2965 T4725 T4724 compound PGKneo,cassette
R2966 T4726 T4713 prep into,inserted
R2967 T4727 T4726 pobj exon,into
R2968 T4728 T4727 nummod 8,exon
R2969 T4729 T4727 prep of,exon
R2970 T4730 T4731 det the,gene
R2971 T4731 T4729 pobj gene,of
R2972 T4732 T4731 compound Adam22,gene
R2973 T4733 T4713 punct .,inserted
R2974 T4735 T4736 det The,construct
R2975 T4736 T4739 nsubj construct,consisted
R2976 T4737 T4736 amod final,construct
R2977 T4738 T4736 compound targeting,construct
R2978 T4740 T4739 prep of,consisted
R2979 T4741 T4742 det a,fragment
R2980 T4742 T4740 pobj fragment,of
R2981 T4743 T4744 nummod 10.5,kb
R2982 T4744 T4742 nmod kb,fragment
R2983 T4745 T4744 punct -,kb
R2984 T4746 T4747 amod genomic,DNA
R2985 T4747 T4742 compound DNA,fragment
R2986 T4748 T4749 dep that,interrupted
R2987 T4749 T4742 relcl interrupted,fragment
R2988 T4750 T4749 auxpass was,interrupted
R2989 T4751 T4749 agent by,interrupted
R2990 T4752 T4753 det the,insertion
R2991 T4753 T4751 pobj insertion,by
R2992 T4754 T4753 prep of,insertion
R2993 T4755 T4756 det the,cassette
R2994 T4756 T4754 pobj cassette,of
R2995 T4757 T4756 compound PGKneo,cassette
R2996 T4758 T4749 cc and,interrupted
R2997 T4759 T4749 conj contained,interrupted
R2998 T4760 T4761 det a,cassette
R2999 T4761 T4759 dobj cassette,contained
R3000 T4762 T4763 compound MC1,TK
R3001 T4763 T4761 compound TK,cassette
R3002 T4764 T4763 punct /,TK
R3003 T4765 T4759 prep as,contained
R3004 T4766 T4767 det a,marker
R3005 T4767 T4765 pobj marker,as
R3006 T4768 T4767 amod negative,marker
R3007 T4769 T4767 compound selection,marker
R3008 T4770 T4767 prep against,marker
R3009 T4771 T4772 amod random,integration
R3010 T4772 T4770 pobj integration,against
R3011 T4773 T4774 punct [,5
R3012 T4774 T4739 parataxis 5,consisted
R3013 T4775 T4774 punct ],5
R3014 T4776 T4739 punct .,consisted
R3019 T4852 T4851 prep of,Generation
R3020 T4853 T4854 npadvmod Adam22,deficient
R3021 T4854 T4855 amod deficient,mice
R3022 T4855 T4852 pobj mice,of
R3023 T4857 T4858 det The,vector
R3024 T4858 T4861 nsubjpass vector,electroporated
R3025 T4859 T4858 amod linearised,vector
R3026 T4860 T4858 compound targeting,vector
R3027 T4862 T4861 auxpass was,electroporated
R3028 T4863 T4861 prep into,electroporated
R3029 T4864 T4865 nmod TT2,cells
R3030 T4865 T4863 pobj cells,into
R3031 T4866 T4867 amod embryonic,stem
R3032 T4867 T4865 nmod stem,cells
R3033 T4868 T4867 punct (,stem
R3034 T4869 T4867 appos ES,stem
R3035 T4870 T4865 punct ),cells
R3036 T4871 T4872 punct [,31
R3037 T4872 T4861 parataxis 31,electroporated
R3038 T4873 T4872 punct ],31
R3039 T4874 T4861 punct .,electroporated
R3040 T4876 T4877 amod Homologous,recombinants
R3041 T4877 T4878 nsubjpass recombinants,selected
R3042 T4879 T4878 auxpass were,selected
R3043 T4880 T4878 prep by,selected
R3044 T4881 T4880 pobj PCR,by
R3045 T4882 T4878 punct .,selected
R3046 T4884 T4885 det The,DNA
R3047 T4885 T4888 nsubjpass DNA,amplified
R3048 T4886 T4885 amod extracted,DNA
R3049 T4887 T4885 amod genomic,DNA
R3050 T4889 T4885 prep from,DNA
R3051 T4890 T4891 det each,clone
R3052 T4891 T4889 pobj clone,from
R3053 T4892 T4888 auxpass was,amplified
R3054 T4893 T4888 advcl using,amplified
R3055 T4894 T4895 det the,primer
R3056 T4895 T4893 dobj primer,using
R3057 T4896 T4895 amod forward,primer
R3058 T4897 T4895 appos AGN2,primer
R3059 T4898 T4895 punct : ,primer
R3060 T4899 T4900 nummod 5,GCCTGCTTGCCGAATATCATGGTGGAAAAT
R3061 T4900 T4895 appos GCCTGCTTGCCGAATATCATGGTGGAAAAT,primer
R3062 T4901 T4899 punct ',5
R3063 T4902 T4900 punct -,GCCTGCTTGCCGAATATCATGGTGGAAAAT
R3064 T4903 T4900 punct -,GCCTGCTTGCCGAATATCATGGTGGAAAAT
R3065 T4904 T4900 nummod 3,GCCTGCTTGCCGAATATCATGGTGGAAAAT
R3066 T4905 T4900 punct ',GCCTGCTTGCCGAATATCATGGTGGAAAAT
R3067 T4906 T4895 prep in,primer
R3068 T4907 T4908 det the,cassette
R3069 T4908 T4906 pobj cassette,in
R3070 T4909 T4908 compound PGKneo,cassette
R3071 T4910 T4895 punct ", ",primer
R3072 T4911 T4895 cc and,primer
R3073 T4912 T4913 det the,primer
R3074 T4913 T4895 conj primer,primer
R3075 T4914 T4913 amod reverse,primer
R3076 T4915 T4913 appos MFP065R,primer
R3077 T4916 T4913 punct : ,primer
R3078 T4917 T4918 nummod 5,ACTATTTCTGTGATGAGGGCACAGCATC
R3079 T4918 T4913 appos ACTATTTCTGTGATGAGGGCACAGCATC,primer
R3080 T4919 T4917 punct ',5
R3081 T4920 T4918 punct -,ACTATTTCTGTGATGAGGGCACAGCATC
R3082 T4921 T4918 punct -,ACTATTTCTGTGATGAGGGCACAGCATC
R3083 T4922 T4918 nummod 3,ACTATTTCTGTGATGAGGGCACAGCATC
R3084 T4923 T4918 punct ',ACTATTTCTGTGATGAGGGCACAGCATC
R3085 T4924 T4913 prep outside,primer
R3086 T4925 T4926 det the,vector
R3087 T4926 T4924 pobj vector,outside
R3088 T4927 T4926 compound targeting,vector
R3089 T4928 T4888 punct .,amplified
R3090 T4930 T4931 amod Homologous,DNA
R3091 T4931 T4933 nsubjpass DNA,amplified
R3092 T4932 T4931 amod recombined,DNA
R3093 T4934 T4933 auxpass was,amplified
R3094 T4935 T4933 advmod efficiently,amplified
R3095 T4936 T4933 prep by,amplified
R3096 T4937 T4938 nummod 35,cycles
R3097 T4938 T4936 pobj cycles,by
R3098 T4939 T4938 prep of,cycles
R3099 T4940 T4941 det the,steps
R3100 T4941 T4939 pobj steps,of
R3101 T4942 T4941 amod following,steps
R3102 T4943 T4941 compound amplification,steps
R3103 T4944 T4941 punct (,steps
R3104 T4945 T4946 nummod 94,°C
R3105 T4946 T4941 appos °C,steps
R3106 T4947 T4946 punct -,°C
R3107 T4948 T4949 nummod 30,s
R3108 T4949 T4946 npadvmod s,°C
R3109 T4950 T4946 cc and,°C
R3110 T4951 T4952 nummod 68,°C
R3111 T4952 T4946 conj °C,°C
R3112 T4953 T4952 punct -,°C
R3113 T4954 T4955 nummod 5,min
R3114 T4955 T4952 npadvmod min,°C
R3115 T4956 T4933 punct ),amplified
R3116 T4957 T4933 punct .,amplified
R3117 T4959 T4960 det The,efficiency
R3118 T4960 T4962 nsubj efficiency,was
R3119 T4961 T4960 compound targeting,efficiency
R3120 T4963 T4960 prep of,efficiency
R3121 T4964 T4965 det this,construct
R3122 T4965 T4963 pobj construct,of
R3123 T4966 T4967 advmod about,4
R3124 T4967 T4968 nummod 4,%
R3125 T4968 T4962 attr %,was
R3126 T4969 T4962 punct .,was
R3127 T4971 T4972 advmod Correctly,targeted
R3128 T4972 T4973 amod targeted,clones
R3129 T4973 T4975 nsubjpass clones,injected
R3130 T4974 T4973 compound ES,clones
R3131 T4976 T4975 auxpass were,injected
R3132 T4977 T4975 prep into,injected
R3133 T4978 T4979 amod fertilized,eggs
R3134 T4979 T4977 pobj eggs,into
R3135 T4980 T4979 compound ICR,eggs
R3136 T4981 T4979 compound mouse,eggs
R3137 T4982 T4975 prep at,injected
R3138 T4983 T4984 det the,stage
R3139 T4984 T4982 pobj stage,at
R3140 T4985 T4986 nummod eight,cell
R3141 T4986 T4984 compound cell,stage
R3142 T4987 T4986 punct -,cell
R3143 T4988 T4975 punct .,injected
R3144 T4990 T4991 det The,chimeras
R3145 T4991 T4994 nsubjpass chimeras,mated
R3146 T4992 T4991 amod resulting,chimeras
R3147 T4993 T4991 amod male,chimeras
R3148 T4995 T4994 auxpass were,mated
R3149 T4996 T4994 prep with,mated
R3150 T4997 T4998 nmod C57BL,females
R3151 T4998 T4996 pobj females,with
R3152 T4999 T4997 punct /,C57BL
R3153 T5000 T4997 nummod 6N,C57BL
R3154 T5001 T4994 punct ", ",mated
R3155 T5002 T4994 cc and,mated
R3156 T5003 T5004 amod heterozygous,mice
R3157 T5004 T5008 nsubjpass mice,interbred
R3158 T5005 T5004 amod male,mice
R3159 T5006 T5005 cc and,male
R3160 T5007 T5005 conj female,male
R3161 T5008 T4994 conj interbred,mated
R3162 T5009 T5008 auxpass were,interbred
R3163 T5010 T5011 aux to,generate
R3164 T5011 T5008 advcl generate,interbred
R3165 T5012 T5013 amod homozygous,mice
R3166 T5013 T5011 dobj mice,generate
R3167 T5014 T5008 punct .,interbred
R3168 T5016 T5017 det The,phenotypes
R3169 T5017 T5019 nsubjpass phenotypes,observed
R3170 T5018 T5017 amod ataxic,phenotypes
R3171 T5020 T5017 prep of,phenotypes
R3172 T5021 T5022 amod homozygous,mice
R3173 T5022 T5020 pobj mice,of
R3174 T5023 T5019 auxpass were,observed
R3175 T5024 T5019 prep in,observed
R3176 T5025 T5026 nummod two,lines
R3177 T5026 T5024 pobj lines,in
R3178 T5027 T5026 amod independent,lines
R3179 T5028 T5019 punct .,observed
R3180 T5055 T5056 compound Southern,blot
R3181 T5056 T5057 compound blot,analysis
R3182 T5059 T5060 nmod Mouse,DNA
R3183 T5060 T5062 nsubjpass DNA,extracted
R3184 T5061 T5060 amod genomic,DNA
R3185 T5063 T5060 acl used,DNA
R3186 T5064 T5063 prep for,used
R3187 T5065 T5066 compound Southern,blot
R3188 T5066 T5067 compound blot,analysis
R3189 T5067 T5064 pobj analysis,for
R3190 T5068 T5062 auxpass was,extracted
R3191 T5069 T5062 prep from,extracted
R3192 T5070 T5071 det the,liver
R3193 T5071 T5069 pobj liver,from
R3194 T5072 T5071 prep of,liver
R3195 T5073 T5074 det each,genotype
R3196 T5074 T5072 pobj genotype,of
R3197 T5075 T5062 punct .,extracted
R3198 T5077 T5078 npadvmod BamHI,digested
R3199 T5078 T5080 amod digested,DNA
R3200 T5079 T5078 punct -,digested
R3201 T5080 T5082 nsubjpass DNA,probed
R3202 T5081 T5080 amod genomic,DNA
R3203 T5083 T5082 auxpass was,probed
R3204 T5084 T5082 prep with,probed
R3205 T5085 T5086 det the,fragment
R3206 T5086 T5084 pobj fragment,with
R3207 T5087 T5086 punct [,fragment
R3208 T5088 T5089 advmod 32P,labelled
R3209 T5089 T5086 amod labelled,fragment
R3210 T5090 T5089 punct ],labelled
R3211 T5091 T5089 punct -,labelled
R3212 T5092 T5093 nummod 0.4,kb
R3213 T5093 T5086 nmod kb,fragment
R3214 T5094 T5093 punct -,kb
R3215 T5095 T5096 nmod SpeI,BamHI
R3216 T5096 T5086 nmod BamHI,fragment
R3217 T5097 T5096 punct -,BamHI
R3218 T5098 T5086 amod genomic,fragment
R3219 T5099 T5086 punct ", ",fragment
R3220 T5100 T5101 dep which,located
R3221 T5101 T5086 relcl located,fragment
R3222 T5102 T5101 auxpass is,located
R3223 T5103 T5101 prep between,located
R3224 T5104 T5105 nmod exons,8
R3225 T5105 T5103 pobj 8,between
R3226 T5106 T5105 cc and,8
R3227 T5107 T5105 conj 9,8
R3228 T5108 T5082 punct .,probed
R3229 T5223 T5224 compound Antibody,production
R3230 T5226 T5227 npadvmod His,tagged
R3231 T5227 T5229 amod tagged,protein
R3232 T5228 T5227 punct -,tagged
R3233 T5229 T5235 nsubjpass protein,produced
R3234 T5230 T5227 cc and,tagged
R3235 T5231 T5232 npadvmod MBP,fused
R3236 T5232 T5227 conj fused,tagged
R3237 T5233 T5232 punct -,fused
R3238 T5234 T5229 amod recombinant,protein
R3239 T5236 T5229 acl containing,protein
R3240 T5237 T5238 amod cytoplasmic,domain
R3241 T5238 T5236 dobj domain,containing
R3242 T5239 T5238 punct (,domain
R3243 T5240 T5238 appos cp,domain
R3244 T5241 T5238 punct ),domain
R3245 T5242 T5238 prep of,domain
R3246 T5243 T5244 amod human,isoform
R3247 T5244 T5242 pobj isoform,of
R3248 T5245 T5244 compound ADAM22,isoform
R3249 T5246 T5244 nummod 1,isoform
R3250 T5247 T5248 punct (,acids
R3251 T5248 T5244 parataxis acids,isoform
R3252 T5249 T5248 dep position,acids
R3253 T5250 T5249 punct : ,position
R3254 T5251 T5252 nummod 756,906
R3255 T5252 T5249 appos 906,position
R3256 T5253 T5252 punct –,906
R3257 T5254 T5248 punct ", ",acids
R3258 T5255 T5248 nummod 151,acids
R3259 T5256 T5248 compound amino,acids
R3260 T5257 T5248 prep in,acids
R3261 T5258 T5257 pobj length,in
R3262 T5259 T5248 punct ),acids
R3263 T5260 T5235 auxpass were,produced
R3264 T5261 T5235 prep in,produced
R3265 T5262 T5263 compound E.,coli
R3266 T5263 T5261 pobj coli,in
R3267 T5264 T5235 punct .,produced
R3268 T5266 T5267 npadvmod His,tagged
R3269 T5267 T5269 amod tagged,protein
R3270 T5268 T5267 punct -,tagged
R3271 T5269 T5273 nsubjpass protein,purified
R3272 T5270 T5271 compound ADAM22,cp
R3273 T5271 T5269 compound cp,protein
R3274 T5272 T5271 punct -,cp
R3275 T5274 T5273 auxpass was,purified
R3276 T5275 T5273 prep in,purified
R3277 T5276 T5277 amod denatured,condition
R3278 T5277 T5275 pobj condition,in
R3279 T5278 T5273 advcl using,purified
R3280 T5279 T5280 compound Ni,NTA
R3281 T5280 T5282 compound NTA,resin
R3282 T5281 T5280 punct -,NTA
R3283 T5282 T5278 dobj resin,using
R3284 T5283 T5284 punct (,GmbH
R3285 T5284 T5282 parataxis GmbH,resin
R3286 T5285 T5284 compound Qiagen,GmbH
R3287 T5286 T5284 punct ),GmbH
R3288 T5287 T5273 cc and,purified
R3289 T5288 T5273 conj dialysed,purified
R3290 T5289 T5288 prep in,dialysed
R3291 T5290 T5289 pobj PBS,in
R3292 T5291 T5273 punct .,purified
R3293 T5293 T5294 amod Precipitated,protein
R3294 T5294 T5295 nsubjpass protein,recovered
R3295 T5295 T5297 ccomp recovered,used
R3296 T5296 T5295 auxpass was,recovered
R3297 T5298 T5295 cc and,recovered
R3298 T5299 T5300 auxpass was,mixed
R3299 T5300 T5295 conj mixed,recovered
R3300 T5301 T5300 prep with,mixed
R3301 T5302 T5303 poss Freund,adjuvant
R3302 T5303 T5301 pobj adjuvant,with
R3303 T5304 T5302 case 's,Freund
R3304 T5305 T5303 amod complete,adjuvant
R3305 T5306 T5307 punct (,Invitrogen
R3306 T5307 T5303 parataxis Invitrogen,adjuvant
R3307 T5308 T5307 punct ),Invitrogen
R3308 T5309 T5297 punct ", ",used
R3309 T5310 T5297 advmod then,used
R3310 T5311 T5297 punct ", ",used
R3311 T5312 T5313 det the,mixture
R3312 T5313 T5297 nsubjpass mixture,used
R3313 T5314 T5297 auxpass was,used
R3314 T5315 T5297 prep for,used
R3315 T5316 T5315 pobj immunization,for
R3316 T5317 T5316 prep of,immunization
R3317 T5318 T5317 pobj rabbits,of
R3318 T5319 T5297 punct .,used
R3319 T5321 T5322 det The,antiserum
R3320 T5322 T5323 nsubjpass antiserum,incubated
R3321 T5324 T5322 acl raised,antiserum
R3322 T5325 T5324 prep against,raised
R3323 T5326 T5327 det the,protein
R3324 T5327 T5325 pobj protein,against
R3325 T5328 T5329 npadvmod His,tagged
R3326 T5329 T5327 amod tagged,protein
R3327 T5330 T5329 punct -,tagged
R3328 T5331 T5332 compound ADAM22,cp
R3329 T5332 T5327 compound cp,protein
R3330 T5333 T5332 punct -,cp
R3331 T5334 T5323 auxpass was,incubated
R3332 T5335 T5323 prep with,incubated
R3333 T5336 T5337 npadvmod MBP,fused
R3334 T5337 T5339 amod fused,protein
R3335 T5338 T5337 punct -,fused
R3336 T5339 T5335 pobj protein,with
R3337 T5340 T5341 compound ADAM22,cp
R3338 T5341 T5339 compound cp,protein
R3339 T5342 T5341 punct -,cp
R3340 T5343 T5339 acl coupled,protein
R3341 T5344 T5343 prep to,coupled
R3342 T5345 T5346 nmod Affi,Gel
R3343 T5346 T5348 nmod Gel,beads
R3344 T5347 T5346 punct -,Gel
R3345 T5348 T5344 pobj beads,to
R3346 T5349 T5346 nummod 10,Gel
R3347 T5350 T5351 punct (,Rad
R3348 T5351 T5348 parataxis Rad,beads
R3349 T5352 T5351 compound Bio,Rad
R3350 T5353 T5351 punct -,Rad
R3351 T5354 T5351 punct ),Rad
R3352 T5355 T5323 punct .,incubated
R3353 T5357 T5358 nsubjpass Beads,washed
R3354 T5359 T5357 prep with,Beads
R3355 T5360 T5361 amod bound,antibodies
R3356 T5361 T5359 pobj antibodies,with
R3357 T5362 T5358 auxpass were,washed
R3358 T5363 T5358 prep in,washed
R3359 T5364 T5363 pobj PBS,in
R3360 T5365 T5358 punct ", ",washed
R3361 T5366 T5358 cc and,washed
R3362 T5367 T5368 det the,antibodies
R3363 T5368 T5370 nsubjpass antibodies,eluted
R3364 T5369 T5368 amod bound,antibodies
R3365 T5370 T5358 conj eluted,washed
R3366 T5371 T5370 auxpass were,eluted
R3367 T5372 T5370 prep with,eluted
R3368 T5373 T5374 nummod 100,mM
R3369 T5374 T5375 compound mM,glycine
R3370 T5375 T5372 pobj glycine,with
R3371 T5376 T5375 punct ", ",glycine
R3372 T5377 T5375 npadvmod pH,glycine
R3373 T5378 T5377 nummod 3.0,pH
R3374 T5379 T5370 punct ", ",eluted
R3375 T5380 T5370 cc and,eluted
R3376 T5381 T5370 conj neutralized,eluted
R3377 T5382 T5381 advmod immediately,neutralized
R3378 T5383 T5370 punct .,eluted
R3379 T5385 T5386 advcl Using,detected
R3380 T5387 T5388 det the,antibody
R3381 T5388 T5385 dobj antibody,Using
R3382 T5389 T5390 npadvmod affinity,purified
R3383 T5390 T5388 amod purified,antibody
R3384 T5391 T5386 punct ", ",detected
R3385 T5392 T5393 preconj both,mouse
R3386 T5393 T5396 compound mouse,proteins
R3387 T5394 T5393 amod human,mouse
R3388 T5395 T5393 cc and,mouse
R3389 T5396 T5386 nsubjpass proteins,detected
R3390 T5397 T5396 compound ADAM22,proteins
R3391 T5398 T5386 auxpass were,detected
R3392 T5399 T5386 prep by,detected
R3393 T5400 T5401 compound Western,blot
R3394 T5401 T5402 compound blot,analysis
R3395 T5402 T5399 pobj analysis,by
R3396 T5403 T5386 punct .,detected
R3397 T5405 T5406 advmod However,was
R3398 T5407 T5406 punct ", ",was
R3399 T5408 T5406 advmod unfortunately,was
R3400 T5409 T5410 det the,antibody
R3401 T5410 T5406 nsubj antibody,was
R3402 T5411 T5406 neg not,was
R3403 T5412 T5406 acomp suitable,was
R3404 T5413 T5412 prep for,suitable
R3405 T5414 T5415 amod immunohistochemical,analysis
R3406 T5415 T5413 pobj analysis,for
R3407 T5416 T5415 prep of,analysis
R3408 T5417 T5418 compound mouse,tissues
R3409 T5418 T5416 pobj tissues,of
R3410 T5419 T5406 punct .,was
R3411 T5499 T5500 compound Western,blot
R3412 T5500 T5501 compound blot,analysis
R3413 T5503 T5504 det The,absence
R3414 T5504 T5505 nsubjpass absence,confirmed
R3415 T5506 T5504 prep of,absence
R3416 T5507 T5508 compound ADAM22,protein
R3417 T5508 T5506 pobj protein,of
R3418 T5509 T5504 prep in,absence
R3419 T5510 T5511 det the,mice
R3420 T5511 T5509 pobj mice,in
R3421 T5512 T5511 nmod Adam22,mice
R3422 T5513 T5512 punct -,Adam22
R3423 T5514 T5512 punct /,Adam22
R3424 T5515 T5512 punct -,Adam22
R3425 T5516 T5505 auxpass was,confirmed
R3426 T5517 T5505 prep by,confirmed
R3427 T5518 T5519 compound Western,blot
R3428 T5519 T5520 compound blot,analysis
R3429 T5520 T5517 pobj analysis,by
R3430 T5521 T5505 punct .,confirmed
R3431 T5523 T5524 advmod Briefly,isolated
R3432 T5525 T5524 punct ", ",isolated
R3433 T5526 T5527 det the,cerebellum
R3434 T5527 T5524 nsubjpass cerebellum,isolated
R3435 T5528 T5524 auxpass was,isolated
R3436 T5529 T5524 prep from,isolated
R3437 T5530 T5531 det a,mouse
R3438 T5531 T5529 pobj mouse,from
R3439 T5532 T5531 compound P14.5,mouse
R3440 T5533 T5531 prep of,mouse
R3441 T5534 T5535 det each,genotype
R3442 T5535 T5533 pobj genotype,of
R3443 T5536 T5524 cc and,isolated
R3444 T5537 T5524 conj homogenized,isolated
R3445 T5538 T5537 prep with,homogenized
R3446 T5539 T5540 det a,homogenizer
R3447 T5540 T5538 pobj homogenizer,with
R3448 T5541 T5540 compound Polytron,homogenizer
R3449 T5542 T5540 prep in,homogenizer
R3450 T5543 T5544 compound cell,lysis
R3451 T5544 T5545 compound lysis,buffer
R3452 T5545 T5542 pobj buffer,in
R3453 T5546 T5547 punct (,Diagnostics
R3454 T5547 T5545 parataxis Diagnostics,buffer
R3455 T5548 T5549 nummod 50,mM
R3456 T5549 T5550 compound mM,HCl
R3457 T5550 T5547 dep HCl,Diagnostics
R3458 T5551 T5550 compound Tris,HCl
R3459 T5552 T5550 punct -,HCl
R3460 T5553 T5550 punct ", ",HCl
R3461 T5554 T5550 npadvmod pH,HCl
R3462 T5555 T5554 nummod 7.5,pH
R3463 T5556 T5550 punct ", ",HCl
R3464 T5557 T5558 nummod 100,mM
R3465 T5558 T5559 compound mM,NaCl
R3466 T5559 T5550 appos NaCl,HCl
R3467 T5560 T5550 punct ", ",HCl
R3468 T5561 T5562 nummod 1,%
R3469 T5562 T5563 compound %,NP
R3470 T5563 T5550 appos NP,HCl
R3471 T5564 T5563 punct -,NP
R3472 T5565 T5563 nummod 40,NP
R3473 T5566 T5550 punct ", ",HCl
R3474 T5567 T5568 amod Complete,inhibitors
R3475 T5568 T5550 appos inhibitors,HCl
R3476 T5569 T5568 compound protease,inhibitors
R3477 T5570 T5547 punct [,Diagnostics
R3478 T5571 T5547 compound Roche,Diagnostics
R3479 T5572 T5547 punct ],Diagnostics
R3480 T5573 T5547 punct ),Diagnostics
R3481 T5574 T5524 punct .,isolated
R3482 T5576 T5577 prep After,separated
R3483 T5578 T5576 pobj removal,After
R3484 T5579 T5578 prep of,removal
R3485 T5580 T5581 compound cell,debris
R3486 T5581 T5579 pobj debris,of
R3487 T5582 T5578 prep by,removal
R3488 T5583 T5582 pobj centrifugation,by
R3489 T5584 T5577 punct ", ",separated
R3490 T5585 T5586 det the,supernatant
R3491 T5586 T5577 nsubjpass supernatant,separated
R3492 T5587 T5577 auxpass was,separated
R3493 T5588 T5577 prep on,separated
R3494 T5589 T5590 nummod 10,%
R3495 T5590 T5591 compound %,PAGE
R3496 T5591 T5588 pobj PAGE,on
R3497 T5592 T5591 compound SDS,PAGE
R3498 T5593 T5591 punct -,PAGE
R3499 T5594 T5577 punct ", ",separated
R3500 T5595 T5577 cc and,separated
R3501 T5596 T5577 conj transferred,separated
R3502 T5597 T5596 prep to,transferred
R3503 T5598 T5599 det a,membrane
R3504 T5599 T5597 pobj membrane,to
R3505 T5600 T5599 compound nitrocellulose,membrane
R3506 T5601 T5577 punct .,separated
R3507 T5603 T5604 det The,blot
R3508 T5604 T5605 nsubjpass blot,incubated
R3509 T5606 T5605 auxpass was,incubated
R3510 T5607 T5605 advmod then,incubated
R3511 T5608 T5605 prep with,incubated
R3512 T5609 T5610 amod polyclonal,antibody
R3513 T5610 T5608 pobj antibody,with
R3514 T5611 T5610 prep against,antibody
R3515 T5612 T5613 det the,domain
R3516 T5613 T5611 pobj domain,against
R3517 T5614 T5613 amod cytoplasmic,domain
R3518 T5615 T5613 prep of,domain
R3519 T5616 T5615 pobj ADAM22,of
R3520 T5617 T5610 punct (,antibody
R3521 T5618 T5610 prep at,antibody
R3522 T5619 T5618 pobj 1,at
R3523 T5620 T5621 punct :,"1,000"
R3524 T5621 T5619 prep "1,000",1
R3525 T5622 T5605 punct ),incubated
R3526 T5623 T5605 punct .,incubated
R3527 T5625 T5626 amod Bound,antibodies
R3528 T5626 T5627 nsubjpass antibodies,visualized
R3529 T5628 T5627 auxpass were,visualized
R3530 T5629 T5627 prep with,visualized
R3531 T5630 T5631 nmod horseradish,antibody
R3532 T5631 T5629 pobj antibody,with
R3533 T5632 T5633 npadvmod peroxidase,labelled
R3534 T5633 T5631 amod labelled,antibody
R3535 T5634 T5633 punct -,labelled
R3536 T5635 T5631 amod second,antibody
R3537 T5636 T5631 cc and,antibody
R3538 T5637 T5638 det a,system
R3539 T5638 T5631 conj system,antibody
R3540 T5639 T5640 compound ECL,plus
R3541 T5640 T5638 compound plus,system
R3542 T5641 T5640 punct -,plus
R3543 T5642 T5638 compound chemiluminescence,system
R3544 T5643 T5638 compound detection,system
R3545 T5644 T5645 punct (,Corp.
R3546 T5645 T5638 parataxis Corp.,system
R3547 T5646 T5645 compound Amersham,Corp.
R3548 T5647 T5645 compound Biosciences,Corp.
R3549 T5648 T5645 punct ),Corp.
R3550 T5649 T5627 punct .,visualized
R3557 T5703 T5704 amod Primary,cell
R3558 T5704 T5706 compound cell,culture
R3559 T5705 T5704 compound Schwann,cell
R3560 T5708 T5709 amod Primary,cells
R3561 T5709 T5711 nsubjpass cells,prepared
R3562 T5710 T5709 compound Schwann,cells
R3563 T5712 T5711 auxpass were,prepared
R3564 T5713 T5711 prep from,prepared
R3565 T5714 T5715 nummod 4,month
R3566 T5715 T5717 npadvmod month,old
R3567 T5716 T5715 punct -,month
R3568 T5717 T5719 amod old,mice
R3569 T5718 T5717 punct -,old
R3570 T5719 T5713 pobj mice,from
R3571 T5720 T5719 nmod C57BL,mice
R3572 T5721 T5720 punct /,C57BL
R3573 T5722 T5720 nummod 6,C57BL
R3574 T5723 T5711 prep according,prepared
R3575 T5724 T5723 prep to,according
R3576 T5725 T5726 poss Manent,protocol
R3577 T5726 T5724 pobj protocol,to
R3578 T5727 T5725 case 's,Manent
R3579 T5728 T5729 punct [,32
R3580 T5729 T5726 parataxis 32,protocol
R3581 T5730 T5729 punct ],32
R3582 T5731 T5726 prep with,protocol
R3583 T5732 T5733 amod minor,modifications
R3584 T5733 T5731 pobj modifications,with
R3585 T5734 T5711 punct .,prepared
R3586 T5736 T5737 advmod Briefly,removed
R3587 T5738 T5737 punct ", ",removed
R3588 T5739 T5740 amod sciatic,nerves
R3589 T5740 T5737 nsubjpass nerves,removed
R3590 T5741 T5737 auxpass were,removed
R3591 T5742 T5737 cc and,removed
R3592 T5743 T5737 conj incubated,removed
R3593 T5744 T5743 prep for,incubated
R3594 T5745 T5746 nummod 7,days
R3595 T5746 T5744 pobj days,for
R3596 T5747 T5743 prep in,incubated
R3597 T5748 T5749 det the,medium
R3598 T5749 T5747 pobj medium,in
R3599 T5750 T5749 amod pre-treatment,medium
R3600 T5751 T5749 punct ", ",medium
R3601 T5752 T5753 dep which,consisted
R3602 T5753 T5749 relcl consisted,medium
R3603 T5754 T5753 prep of,consisted
R3604 T5755 T5756 compound D,MEM
R3605 T5756 T5754 pobj MEM,of
R3606 T5757 T5756 punct -,MEM
R3607 T5758 T5759 punct (,glucose
R3608 T5759 T5756 parataxis glucose,MEM
R3609 T5760 T5759 amod high,glucose
R3610 T5761 T5759 punct ),glucose
R3611 T5762 T5756 acl supplemented,MEM
R3612 T5763 T5762 prep with,supplemented
R3613 T5764 T5765 nummod 10,%
R3614 T5765 T5766 compound %,FCS
R3615 T5766 T5763 pobj FCS,with
R3616 T5767 T5766 punct ", ",FCS
R3617 T5768 T5769 nummod 50,mg
R3618 T5769 T5770 nmod mg,gentamicin
R3619 T5770 T5766 conj gentamicin,FCS
R3620 T5771 T5772 punct /,ml
R3621 T5772 T5769 prep ml,mg
R3622 T5773 T5774 punct (,Corp.
R3623 T5774 T5770 parataxis Corp.,gentamicin
R3624 T5775 T5774 compound Invitrogen,Corp.
R3625 T5776 T5774 punct ),Corp.
R3626 T5777 T5770 punct ", ",gentamicin
R3627 T5778 T5779 nummod 2.5,μg
R3628 T5779 T5780 nmod μg,fungizone
R3629 T5780 T5770 conj fungizone,gentamicin
R3630 T5781 T5782 punct /,ml
R3631 T5782 T5779 prep ml,μg
R3632 T5783 T5784 punct (,Corp.
R3633 T5784 T5780 parataxis Corp.,fungizone
R3634 T5785 T5784 compound Invitrogen,Corp.
R3635 T5786 T5784 punct ),Corp.
R3636 T5787 T5780 punct ", ",fungizone
R3637 T5788 T5789 nummod 2,μM
R3638 T5789 T5790 compound μM,forskolin
R3639 T5790 T5780 conj forskolin,fungizone
R3640 T5791 T5792 punct (,Inc.
R3641 T5792 T5790 parataxis Inc.,forskolin
R3642 T5793 T5792 compound EMD,Inc.
R3643 T5794 T5792 compound Biosciences,Inc.
R3644 T5795 T5792 punct ),Inc.
R3645 T5796 T5790 cc and,forskolin
R3646 T5797 T5798 nummod 10,ng
R3647 T5798 T5799 nmod ng,beta1
R3648 T5799 T5790 conj beta1,forskolin
R3649 T5800 T5801 punct /,ml
R3650 T5801 T5798 prep ml,ng
R3651 T5802 T5799 amod recombinant,beta1
R3652 T5803 T5799 compound heregulin,beta1
R3653 T5804 T5799 punct -,beta1
R3654 T5805 T5806 punct (,Systems
R3655 T5806 T5799 parataxis Systems,beta1
R3656 T5807 T5806 nmod R,Systems
R3657 T5808 T5807 cc &,R
R3658 T5809 T5807 conj D,R
R3659 T5810 T5806 punct ),Systems
R3660 T5811 T5737 punct .,removed
R3661 T5813 T5814 prep For,cut
R3662 T5815 T5813 pobj dissociation,For
R3663 T5816 T5814 punct ", ",cut
R3664 T5817 T5818 amod cultured,nerves
R3665 T5818 T5814 nsubjpass nerves,cut
R3666 T5819 T5818 amod sciatic,nerves
R3667 T5820 T5814 auxpass were,cut
R3668 T5821 T5814 prep into,cut
R3669 T5822 T5821 pobj pieces,into
R3670 T5823 T5814 cc and,cut
R3671 T5824 T5814 conj incubated,cut
R3672 T5825 T5824 prep at,incubated
R3673 T5826 T5827 nummod 37,°C
R3674 T5827 T5825 pobj °C,at
R3675 T5828 T5824 prep for,incubated
R3676 T5829 T5830 nummod 3,hours
R3677 T5830 T5828 pobj hours,for
R3678 T5831 T5824 prep in,incubated
R3679 T5832 T5833 compound Opti,MEM
R3680 T5833 T5835 compound MEM,medium
R3681 T5834 T5833 punct -,MEM
R3682 T5835 T5831 pobj medium,in
R3683 T5836 T5837 punct (,Corp.
R3684 T5837 T5835 parataxis Corp.,medium
R3685 T5838 T5837 compound Invitrogen,Corp.
R3686 T5839 T5837 punct ),Corp.
R3687 T5840 T5835 acl containing,medium
R3688 T5841 T5842 nummod 130,U
R3689 T5842 T5843 nmod U,type
R3690 T5843 T5840 dobj type,containing
R3691 T5844 T5845 punct /,ml
R3692 T5845 T5842 prep ml,U
R3693 T5846 T5843 compound collagenase,type
R3694 T5847 T5843 nummod I,type
R3695 T5848 T5849 punct (,Corp.
R3696 T5849 T5843 parataxis Corp.,type
R3697 T5850 T5849 compound Invitrogen,Corp.
R3698 T5851 T5849 punct ),Corp.
R3699 T5852 T5843 cc and,type
R3700 T5853 T5854 nummod 0.4,U
R3701 T5854 T5855 nmod U,dispase
R3702 T5855 T5843 conj dispase,type
R3703 T5856 T5857 punct /,ml
R3704 T5857 T5854 prep ml,U
R3705 T5858 T5855 nummod II,dispase
R3706 T5859 T5860 punct (,Diagnostics
R3707 T5860 T5855 parataxis Diagnostics,dispase
R3708 T5861 T5860 compound Roche,Diagnostics
R3709 T5862 T5860 punct ),Diagnostics
R3710 T5863 T5814 punct .,cut
R3711 T5865 T5866 amod Dissociated,cells
R3712 T5866 T5867 nsubjpass cells,resuspended
R3713 T5868 T5867 auxpass were,resuspended
R3714 T5869 T5867 prep in,resuspended
R3715 T5870 T5871 det the,medium
R3716 T5871 T5869 pobj medium,in
R3717 T5872 T5871 amod pre-treatment,medium
R3718 T5873 T5867 cc and,resuspended
R3719 T5874 T5867 conj plated,resuspended
R3720 T5875 T5874 prep on,plated
R3721 T5876 T5877 compound Poly,Lysine
R3722 T5877 T5881 npadvmod Lysine,coated
R3723 T5878 T5877 punct -,Lysine
R3724 T5879 T5877 compound D,Lysine
R3725 T5880 T5877 punct -,Lysine
R3726 T5881 T5884 amod coated,plate
R3727 T5882 T5877 punct /,Lysine
R3728 T5883 T5877 appos Laminin,Lysine
R3729 T5884 T5875 pobj plate,on
R3730 T5885 T5886 punct (,Biosciences
R3731 T5886 T5884 parataxis Biosciences,plate
R3732 T5887 T5886 compound BD,Biosciences
R3733 T5888 T5886 punct ),Biosciences
R3734 T5889 T5867 punct .,resuspended
R3735 T5891 T5892 det The,purity
R3736 T5892 T5893 nsubj purity,approached
R3737 T5894 T5892 prep of,purity
R3738 T5895 T5896 det the,cells
R3739 T5896 T5894 pobj cells,of
R3740 T5897 T5896 amod cultured,cells
R3741 T5898 T5896 compound Schwann,cells
R3742 T5899 T5892 punct ", ",purity
R3743 T5900 T5901 mark as,determined
R3744 T5901 T5892 advcl determined,purity
R3745 T5902 T5901 prep by,determined
R3746 T5903 T5904 amod indirect,analysis
R3747 T5904 T5902 pobj analysis,by
R3748 T5905 T5904 compound immunofluorescence,analysis
R3749 T5906 T5893 punct ", ",approached
R3750 T5907 T5908 nummod 90,%
R3751 T5908 T5893 dobj %,approached
R3752 T5909 T5893 punct .,approached
R3755 T5980 T5981 compound RT,PCR
R3756 T5981 T5983 compound PCR,analysis
R3757 T5982 T5981 punct -,PCR
R3758 T5985 T5986 amod Adult,mice
R3759 T5986 T5991 nsubjpass mice,used
R3760 T5987 T5986 nmod C57BL,mice
R3761 T5988 T5987 punct /,C57BL
R3762 T5989 T5987 nummod 6,C57BL
R3763 T5990 T5986 amod male,mice
R3764 T5992 T5991 auxpass were,used
R3765 T5993 T5991 prep in,used
R3766 T5994 T5995 det this,study
R3767 T5995 T5993 pobj study,in
R3768 T5996 T5991 punct .,used
R3769 T5998 T5999 amod Total,RNAs
R3770 T5999 T6000 nsubjpass RNAs,analysed
R3771 T6001 T5999 acl purified,RNAs
R3772 T6002 T6001 prep from,purified
R3773 T6003 T6004 det the,cerebellum
R3774 T6004 T6002 pobj cerebellum,from
R3775 T6005 T6004 punct ", ",cerebellum
R3776 T6006 T6007 amod spinal,cord
R3777 T6007 T6004 conj cord,cerebellum
R3778 T6008 T6007 punct ", ",cord
R3779 T6009 T6010 amod sciatic,nerve
R3780 T6010 T6007 conj nerve,cord
R3781 T6011 T6010 punct ", ",nerve
R3782 T6012 T6010 conj DRG,nerve
R3783 T6013 T6012 cc and,DRG
R3784 T6014 T6015 amod cultured,cells
R3785 T6015 T6012 conj cells,DRG
R3786 T6016 T6015 compound Schwann,cells
R3787 T6017 T6001 advcl using,purified
R3788 T6018 T6017 dobj TRIzol,using
R3789 T6019 T6020 punct (,Corp.
R3790 T6020 T6018 parataxis Corp.,TRIzol
R3791 T6021 T6020 compound Invitrogen,Corp.
R3792 T6022 T6020 punct ),Corp.
R3793 T6023 T6018 cc and,TRIzol
R3794 T6024 T6025 compound RNeasy,kit
R3795 T6025 T6018 conj kit,TRIzol
R3796 T6026 T6027 punct (,GmbH
R3797 T6027 T6025 parataxis GmbH,kit
R3798 T6028 T6027 compound Qiagen,GmbH
R3799 T6029 T6027 punct ),GmbH
R3800 T6030 T6000 auxpass were,analysed
R3801 T6031 T6000 prep by,analysed
R3802 T6032 T6033 compound RT,PCR
R3803 T6033 T6031 pobj PCR,by
R3804 T6034 T6033 punct -,PCR
R3805 T6035 T6000 advcl using,analysed
R3806 T6036 T6035 dobj SuperScript,using
R3807 T6037 T6036 nummod II,SuperScript
R3808 T6038 T6036 cc and,SuperScript
R3809 T6039 T6040 amod random,primer
R3810 T6040 T6036 conj primer,SuperScript
R3811 T6041 T6042 punct (,Corp.
R3812 T6042 T6040 parataxis Corp.,primer
R3813 T6043 T6042 compound Invitrogen,Corp.
R3814 T6044 T6042 punct ),Corp.
R3815 T6045 T6036 punct ", ",SuperScript
R3816 T6046 T6036 cc and,SuperScript
R3817 T6047 T6048 compound PCR,amplification
R3818 T6048 T6036 conj amplification,SuperScript
R3819 T6049 T6050 punct (,°C
R3820 T6050 T6048 parataxis °C,amplification
R3821 T6051 T6052 nummod 40,cycles
R3822 T6052 T6050 dep cycles,°C
R3823 T6053 T6050 punct ;,°C
R3824 T6054 T6050 nummod 94,°C
R3825 T6055 T6050 punct -,°C
R3826 T6056 T6057 nummod 30,s
R3827 T6057 T6050 npadvmod s,°C
R3828 T6058 T6050 punct ", ",°C
R3829 T6059 T6060 nummod 60,°C
R3830 T6060 T6050 conj °C,°C
R3831 T6061 T6060 punct -,°C
R3832 T6062 T6063 nummod 30,s
R3833 T6063 T6060 npadvmod s,°C
R3834 T6064 T6060 cc and,°C
R3835 T6065 T6066 nummod 68,°C
R3836 T6066 T6060 conj °C,°C
R3837 T6067 T6066 punct -,°C
R3838 T6068 T6069 nummod 5,min
R3839 T6069 T6066 npadvmod min,°C
R3840 T6070 T6050 punct ),°C
R3841 T6071 T6048 prep with,amplification
R3842 T6072 T6073 compound Expand,polymerase
R3843 T6073 T6071 pobj polymerase,with
R3844 T6074 T6075 compound Hi,Fidelity
R3845 T6075 T6073 compound Fidelity,polymerase
R3846 T6076 T6075 punct -,Fidelity
R3847 T6077 T6073 compound DNA,polymerase
R3848 T6078 T6079 punct (,Diagnostics
R3849 T6079 T6073 parataxis Diagnostics,polymerase
R3850 T6080 T6079 compound Roche,Diagnostics
R3851 T6081 T6079 punct ),Diagnostics
R3852 T6082 T6073 cc and,polymerase
R3853 T6083 T6084 npadvmod ADAM22,specific
R3854 T6084 T6086 amod specific,primers
R3855 T6085 T6084 punct -,specific
R3856 T6086 T6073 conj primers,polymerase
R3857 T6087 T6000 punct .,analysed
R3858 T6089 T6090 aux To,detect
R3859 T6090 T6091 advcl detect,placed
R3860 T6092 T6093 compound splicing,variants
R3861 T6093 T6090 dobj variants,detect
R3862 T6094 T6090 prep in,detect
R3863 T6095 T6096 det the,domain
R3864 T6096 T6094 pobj domain,in
R3865 T6097 T6096 amod cytoplasmic,domain
R3866 T6098 T6091 punct ", ",placed
R3867 T6099 T6100 det a,primer
R3868 T6100 T6091 nsubjpass primer,placed
R3869 T6101 T6100 amod forward,primer
R3870 T6102 T6091 auxpass was,placed
R3871 T6103 T6104 advmod just,upstream
R3872 T6104 T6091 advmod upstream,placed
R3873 T6105 T6104 prep of,upstream
R3874 T6106 T6107 det the,domain
R3875 T6107 T6105 pobj domain,of
R3876 T6108 T6107 amod transmembrane,domain
R3877 T6109 T6091 cc and,placed
R3878 T6110 T6111 det a,primer
R3879 T6111 T6113 nsubjpass primer,set
R3880 T6112 T6111 amod reverse,primer
R3881 T6113 T6091 conj set,placed
R3882 T6114 T6113 auxpass was,set
R3883 T6115 T6113 prep on,set
R3884 T6116 T6117 det the,exon
R3885 T6117 T6115 pobj exon,on
R3886 T6118 T6117 amod terminating,exon
R3887 T6119 T6091 punct .,placed
R3888 T6121 T6122 det The,primers
R3889 T6122 T6123 nsubj primers,were
R3890 T6124 T6122 acl used,primers
R3891 T6125 T6124 prep in,used
R3892 T6126 T6127 det this,study
R3893 T6127 T6125 pobj study,in
R3894 T6128 T6129 mark as,follows
R3895 T6129 T6123 advcl follows,were
R3896 T6130 T6123 punct .,were
R3897 T6133 T6132 punct -,ISH04
R3898 T6134 T6132 amod forward,ISH04
R3899 T6135 T6132 punct : ,ISH04
R3900 T6136 T6137 nummod 5,AACAGGCACTGGACAGGGGCTGAC
R3901 T6137 T6132 appos AACAGGCACTGGACAGGGGCTGAC,ISH04
R3902 T6138 T6136 punct ',5
R3903 T6139 T6137 punct -,AACAGGCACTGGACAGGGGCTGAC
R3904 T6140 T6137 punct -,AACAGGCACTGGACAGGGGCTGAC
R3905 T6141 T6137 nummod 3,AACAGGCACTGGACAGGGGCTGAC
R3906 T6142 T6137 punct ',AACAGGCACTGGACAGGGGCTGAC
R3907 T6143 T6132 cc and,ISH04
R3908 T6144 T6132 conj ISH04,ISH04
R3909 T6145 T6144 punct -,ISH04
R3910 T6146 T6144 amod reverse,ISH04
R3911 T6147 T6144 punct : ,ISH04
R3912 T6148 T6149 nummod 5,AATGGATGTCTCCCATAGCCTGGC
R3913 T6149 T6144 appos AATGGATGTCTCCCATAGCCTGGC,ISH04
R3914 T6150 T6148 punct ',5
R3915 T6151 T6149 punct -,AATGGATGTCTCCCATAGCCTGGC
R3916 T6152 T6149 punct -,AATGGATGTCTCCCATAGCCTGGC
R3917 T6153 T6149 nummod 3,AATGGATGTCTCCCATAGCCTGGC
R3918 T6154 T6149 punct ',AATGGATGTCTCCCATAGCCTGGC
R3919 T6155 T6144 punct .,ISH04
R3920 T6214 T6215 nsubjpass Mice,anesthetized
R3921 T6216 T6215 auxpass were,anesthetized
R3922 T6217 T6215 prep with,anesthetized
R3923 T6218 T6219 compound ethyl,ether
R3924 T6219 T6217 pobj ether,with
R3925 T6220 T6215 punct .,anesthetized
R3926 T6222 T6223 amod Whole,body
R3927 T6223 T6225 compound body,perfusion
R3928 T6224 T6223 punct -,body
R3929 T6225 T6226 nsubjpass perfusion,performed
R3930 T6227 T6225 prep by,perfusion
R3931 T6228 T6229 nummod 2,%
R3932 T6229 T6230 compound %,solution
R3933 T6230 T6227 pobj solution,by
R3934 T6231 T6232 compound paraformaldehyde,glutaraldehyde
R3935 T6232 T6230 compound glutaraldehyde,solution
R3936 T6233 T6232 punct -,glutaraldehyde
R3937 T6234 T6225 acl followed,perfusion
R3938 T6235 T6234 agent by,followed
R3939 T6236 T6237 npadvmod heparin,included
R3940 T6237 T6239 amod included,saline
R3941 T6238 T6237 punct -,included
R3942 T6239 T6235 pobj saline,by
R3943 T6240 T6226 auxpass was,performed
R3944 T6241 T6226 punct .,performed
R3945 T6243 T6244 amod Sciatic,nerves
R3946 T6244 T6245 nsubjpass nerves,removed
R3947 T6246 T6244 punct ", ",nerves
R3948 T6247 T6248 amod trigeminal,nerves
R3949 T6248 T6244 conj nerves,nerves
R3950 T6249 T6248 punct ", ",nerves
R3951 T6250 T6248 conj brain,nerves
R3952 T6251 T6250 cc and,brain
R3953 T6252 T6253 amod spinal,cord
R3954 T6253 T6250 conj cord,brain
R3955 T6254 T6245 auxpass were,removed
R3956 T6255 T6245 cc and,removed
R3957 T6256 T6245 conj fixed,removed
R3958 T6257 T6256 prep in,fixed
R3959 T6258 T6259 nummod 10,%
R3960 T6259 T6260 nmod %,formalin
R3961 T6260 T6257 pobj formalin,in
R3962 T6261 T6262 amod neutral,buffered
R3963 T6262 T6260 amod buffered,formalin
R3964 T6263 T6262 punct -,buffered
R3965 T6264 T6245 punct .,removed
R3966 T6266 T6267 det The,cord
R3967 T6267 T6269 nsubjpass cord,washed
R3968 T6268 T6267 amod spinal,cord
R3969 T6270 T6267 cc and,cord
R3970 T6271 T6272 amod other,blocks
R3971 T6272 T6267 conj blocks,cord
R3972 T6273 T6272 compound nerve,blocks
R3973 T6274 T6269 auxpass were,washed
R3974 T6275 T6269 cc and,washed
R3975 T6276 T6269 conj postfixed,washed
R3976 T6277 T6276 prep with,postfixed
R3977 T6278 T6279 nummod 2,%
R3978 T6279 T6280 compound %,tetroxide
R3979 T6280 T6277 pobj tetroxide,with
R3980 T6281 T6280 compound osmium,tetroxide
R3981 T6282 T6269 punct ", ",washed
R3982 T6283 T6269 cc and,washed
R3983 T6284 T6285 auxpass were,dehydrated
R3984 T6285 T6269 conj dehydrated,washed
R3985 T6286 T6285 prep in,dehydrated
R3986 T6287 T6286 pobj ethanol,in
R3987 T6288 T6285 cc and,dehydrated
R3988 T6289 T6285 conj equilibrated,dehydrated
R3989 T6290 T6289 prep in,equilibrated
R3990 T6291 T6290 pobj Epon,in
R3991 T6292 T6269 punct .,washed
R3992 T6294 T6295 npadvmod Epon,embedded
R3993 T6295 T6296 amod embedded,sections
R3994 T6296 T6298 nsubjpass sections,stained
R3995 T6297 T6296 amod semithin,sections
R3996 T6299 T6298 auxpass were,stained
R3997 T6300 T6298 prep with,stained
R3998 T6301 T6302 compound toluidine,blue
R3999 T6302 T6300 pobj blue,with
R4000 T6303 T6298 cc and,stained
R4001 T6304 T6305 auxpass were,subjected
R4002 T6305 T6298 conj subjected,stained
R4003 T6306 T6305 prep to,subjected
R4004 T6307 T6308 npadvmod light,microscopic
R4005 T6308 T6309 amod microscopic,examination
R4006 T6309 T6306 pobj examination,to
R4007 T6310 T6298 punct .,stained
R4008 T6312 T6313 prep For,cut
R4009 T6314 T6315 npadvmod electron,microscopic
R4010 T6315 T6316 amod microscopic,analysis
R4011 T6316 T6312 pobj analysis,For
R4012 T6317 T6313 punct ", ",cut
R4013 T6318 T6319 amod thin,sections
R4014 T6319 T6313 nsubjpass sections,cut
R4015 T6320 T6313 auxpass were,cut
R4016 T6321 T6313 advcl using,cut
R4017 T6322 T6323 det an,ultramicrotome
R4018 T6323 T6321 dobj ultramicrotome,using
R4019 T6324 T6313 punct ", ",cut
R4020 T6325 T6313 conj stained,cut
R4021 T6326 T6325 prep with,stained
R4022 T6327 T6328 nummod 1.5,%
R4023 T6328 T6329 compound %,uranylacetate
R4024 T6329 T6326 pobj uranylacetate,with
R4025 T6330 T6325 prep in,stained
R4026 T6331 T6332 nummod 50,%
R4027 T6332 T6333 compound %,ethanol
R4028 T6333 T6330 pobj ethanol,in
R4029 T6334 T6333 cc and,ethanol
R4030 T6335 T6336 nummod 0.8,%
R4031 T6336 T6337 compound %,citrate
R4032 T6337 T6333 conj citrate,ethanol
R4033 T6338 T6337 compound lead,citrate
R4034 T6339 T6325 punct ", ",stained
R4035 T6340 T6325 cc and,stained
R4036 T6341 T6325 conj analysed,stained
R4037 T6342 T6341 advcl using,analysed
R4038 T6343 T6344 compound electron,microscope
R4039 T6344 T6342 dobj microscope,using
R4040 T6345 T6313 punct .,cut
R4041 T6347 T6348 det All,procedures
R4042 T6348 T6349 nsubjpass procedures,conducted
R4043 T6350 T6349 auxpass were,conducted
R4044 T6351 T6349 prep according,conducted
R4045 T6352 T6351 prep to,according
R4046 T6353 T6354 det the,Committee
R4047 T6354 T6358 poss Committee,guidelines
R4048 T6355 T6354 compound Eisai,Committee
R4049 T6356 T6357 compound Animal,Care
R4050 T6357 T6354 compound Care,Committee
R4051 T6358 T6352 pobj guidelines,to
R4052 T6359 T6354 case 's,Committee
R4053 T6360 T6349 punct .,conducted
R4054 T6427 T6428 amod Frozen,sections
R4055 T6428 T6429 nsubjpass sections,rinsed
R4056 T6430 T6429 auxpass were,rinsed
R4057 T6431 T6429 prep in,rinsed
R4058 T6432 T6433 nummod 0.1,%
R4059 T6433 T6431 pobj %,in
R4060 T6434 T6435 compound Triton,X
R4061 T6435 T6433 appos X,%
R4062 T6436 T6435 punct -,X
R4063 T6437 T6435 nummod 100,X
R4064 T6438 T6433 punct /,%
R4065 T6439 T6433 appos PBS,%
R4066 T6440 T6429 prep at,rinsed
R4067 T6441 T6442 compound room,temperature
R4068 T6442 T6440 pobj temperature,at
R4069 T6443 T6429 prep for,rinsed
R4070 T6444 T6445 nummod 1,hour
R4071 T6445 T6443 pobj hour,for
R4072 T6446 T6429 punct ", ",rinsed
R4073 T6447 T6429 cc and,rinsed
R4074 T6448 T6449 auxpass were,blocked
R4075 T6449 T6429 conj blocked,rinsed
R4076 T6450 T6449 prep in,blocked
R4077 T6451 T6452 compound BLOCKACE,solution
R4078 T6452 T6450 pobj solution,in
R4079 T6453 T6454 punct (,nippon
R4080 T6454 T6452 parataxis nippon,solution
R4081 T6455 T6454 compound Dai,nippon
R4082 T6456 T6454 punct -,nippon
R4083 T6457 T6454 punct ),nippon
R4084 T6458 T6429 punct .,rinsed
R4085 T6460 T6461 det The,antibodies
R4086 T6461 T6463 nsubjpass antibodies,incubated
R4087 T6462 T6461 amod following,antibodies
R4088 T6464 T6463 auxpass were,incubated
R4089 T6465 T6463 advmod overnight,incubated
R4090 T6466 T6463 prep in,incubated
R4091 T6467 T6468 nummod 0.1,%
R4092 T6468 T6469 compound %,BLOCKACE
R4093 T6469 T6466 pobj BLOCKACE,in
R4094 T6470 T6463 prep at,incubated
R4095 T6471 T6472 nummod 4,°C
R4096 T6472 T6470 pobj °C,at
R4097 T6473 T6463 punct :,incubated
R4098 T6475 T6476 amod monoclonal,K
R4099 T6477 T6476 nmod mouse,K
R4100 T6478 T6476 amod anti-calbindin,K
R4101 T6479 T6476 nummod 28,K
R4102 T6480 T6481 punct (,200
R4103 T6481 T6476 parataxis 200,K
R4104 T6482 T6481 dep Sigma,200
R4105 T6483 T6481 punct ", ",200
R4106 T6484 T6481 nummod 1,200
R4107 T6485 T6481 punct :,200
R4108 T6486 T6481 punct ),200
R4109 T6487 T6476 punct ;,K
R4110 T6488 T6489 amod monoclonal,mouse
R4111 T6489 T6476 appos mouse,K
R4112 T6490 T6489 amod anti-MBP,mouse
R4113 T6491 T6492 punct (,50
R4114 T6492 T6489 parataxis 50,mouse
R4115 T6493 T6492 dep SMI,50
R4116 T6494 T6493 punct -,SMI
R4117 T6495 T6493 nummod 99,SMI
R4118 T6496 T6492 punct ", ",50
R4119 T6497 T6498 compound Sternberger,Monoclonals
R4120 T6498 T6492 dep Monoclonals,50
R4121 T6499 T6498 amod Inc.,Monoclonals
R4122 T6500 T6492 punct ", ",50
R4123 T6501 T6492 quantmod 1,50
R4124 T6502 T6492 punct :,50
R4125 T6503 T6492 punct ),50
R4126 T6504 T6476 punct ;,K
R4127 T6505 T6506 amod monoclonal,N
R4128 T6506 T6476 appos N,K
R4129 T6507 T6506 nmod mouse,N
R4130 T6508 T6506 amod anti-Neu,N
R4131 T6509 T6510 punct (,100
R4132 T6510 T6506 parataxis 100,N
R4133 T6511 T6510 dep CHEMICON,100
R4134 T6512 T6510 punct ", ",100
R4135 T6513 T6510 quantmod 1,100
R4136 T6514 T6510 punct :,100
R4137 T6515 T6510 punct ),100
R4138 T6516 T6476 punct ;,K
R4139 T6517 T6476 appos rabbit,K
R4140 T6518 T6517 amod anti-S100,rabbit
R4141 T6519 T6517 amod polyclonal,rabbit
R4142 T6520 T6521 punct (,200
R4143 T6521 T6517 parataxis 200,rabbit
R4144 T6522 T6521 dep DAKO,200
R4145 T6523 T6521 punct ", ",200
R4146 T6524 T6521 quantmod 1,200
R4147 T6525 T6521 punct :,200
R4148 T6526 T6521 punct ),200
R4149 T6527 T6476 punct .,K
R4150 T6529 T6530 nsubjpass Nuclei,counterstained
R4151 T6531 T6530 auxpass were,counterstained
R4152 T6532 T6530 prep with,counterstained
R4153 T6533 T6534 nummod 1,μg
R4154 T6534 T6535 nmod μg,DAPI
R4155 T6535 T6532 pobj DAPI,with
R4156 T6536 T6537 punct /,ml
R4157 T6537 T6534 prep ml,μg
R4158 T6538 T6539 punct (,Sigma
R4159 T6539 T6535 parataxis Sigma,DAPI
R4160 T6540 T6539 punct ),Sigma
R4161 T6541 T6530 punct .,counterstained
R4162 T6543 T6544 compound MBP,staining
R4163 T6544 T6545 nsubjpass staining,performed
R4164 T6546 T6545 auxpass was,performed
R4165 T6547 T6545 prep after,performed
R4166 T6548 T6549 compound microwave,retrieval
R4167 T6549 T6547 pobj retrieval,after
R4168 T6550 T6549 compound epitope,retrieval
R4169 T6551 T6549 prep in,retrieval
R4170 T6552 T6553 nummod 10,mmol
R4171 T6553 T6554 nmod mmol,buffer
R4172 T6554 T6551 pobj buffer,in
R4173 T6555 T6556 punct /,L
R4174 T6556 T6553 prep L,mmol
R4175 T6557 T6554 compound citrate,buffer
R4176 T6558 T6554 punct (,buffer
R4177 T6559 T6554 npadvmod pH,buffer
R4178 T6560 T6559 nummod 6,pH
R4179 T6561 T6554 punct ),buffer
R4180 T6562 T6545 punct .,performed
R4181 T6564 T6565 nsubjpass Sections,incubated
R4182 T6566 T6565 auxpass were,incubated
R4183 T6567 T6565 prep for,incubated
R4184 T6568 T6569 nummod 1,hour
R4185 T6569 T6567 pobj hour,for
R4186 T6570 T6565 prep with,incubated
R4187 T6571 T6572 amod secondary,antibodies
R4188 T6572 T6570 pobj antibodies,with
R4189 T6573 T6572 punct : ,antibodies
R4190 T6574 T6575 npadvmod Cy3,labeled
R4191 T6575 T6577 amod labeled,IgG
R4192 T6576 T6575 punct -,labeled
R4193 T6577 T6572 appos IgG,antibodies
R4194 T6578 T6577 nmod donkey,IgG
R4195 T6579 T6577 amod anti-rabbit,IgG
R4196 T6580 T6581 punct (,200
R4197 T6581 T6577 parataxis 200,IgG
R4198 T6582 T6583 compound Jackson,Research
R4199 T6583 T6581 dep Research,200
R4200 T6584 T6583 compound Immuno,Research
R4201 T6585 T6581 punct ", ",200
R4202 T6586 T6581 quantmod 1,200
R4203 T6587 T6581 punct :,200
R4204 T6588 T6581 punct ),200
R4205 T6589 T6577 punct ", ",IgG
R4206 T6590 T6591 npadvmod FITC,labelled
R4207 T6591 T6593 amod labelled,IgG
R4208 T6592 T6591 punct -,labelled
R4209 T6593 T6577 appos IgG,IgG
R4210 T6594 T6593 nmod donkey,IgG
R4211 T6595 T6593 amod anti-mouse,IgG
R4212 T6596 T6597 punct (,200
R4213 T6597 T6593 parataxis 200,IgG
R4214 T6598 T6599 compound Jackson,Research
R4215 T6599 T6597 dep Research,200
R4216 T6600 T6599 compound Immuno,Research
R4217 T6601 T6597 punct ", ",200
R4218 T6602 T6597 quantmod 1,200
R4219 T6603 T6597 punct :,200
R4220 T6604 T6597 punct ),200
R4221 T6605 T6565 punct .,incubated
R4222 T6607 T6608 nsubjpass Sections,photographed
R4223 T6609 T6608 auxpass were,photographed
R4224 T6610 T6608 prep with,photographed
R4225 T6611 T6612 det an,microscope
R4226 T6612 T6610 pobj microscope,with
R4227 T6613 T6612 compound Olympus,microscope
R4228 T6614 T6615 punct (,IX71
R4229 T6615 T6612 parataxis IX71,microscope
R4230 T6616 T6615 compound Olympus,IX71
R4231 T6617 T6615 punct ),IX71
R4232 T6618 T6612 acl equipped,microscope
R4233 T6619 T6618 prep with,equipped
R4234 T6620 T6621 det a,camera
R4235 T6621 T6619 pobj camera,with
R4236 T6622 T6623 amod high,resolution
R4237 T6623 T6621 compound resolution,camera
R4238 T6624 T6623 punct -,resolution
R4239 T6625 T6621 compound CCD,camera
R4240 T6626 T6608 punct .,photographed
R4241 T6692 T6693 nmod RNA,hybridisation
R4242 T6694 T6695 advmod in,situ
R4243 T6695 T6693 amod situ,hybridisation
R4244 T6697 T6698 advmod In,situ
R4245 T6698 T6699 amod situ,hybridisation
R4246 T6699 T6700 nsubjpass hybridisation,carried
R4247 T6701 T6699 prep on,hybridisation
R4248 T6702 T6703 amod frozen,sections
R4249 T6703 T6701 pobj sections,on
R4250 T6704 T6700 auxpass were,carried
R4251 T6705 T6700 prt out,carried
R4252 T6706 T6700 advcl using,carried
R4253 T6707 T6708 advmod 35S,labelled
R4254 T6708 T6710 amod labelled,probes
R4255 T6709 T6708 punct -,labelled
R4256 T6710 T6706 dobj probes,using
R4257 T6711 T6710 compound RNA,probes
R4258 T6712 T6700 punct .,carried
R4259 T6714 T6715 advmod Briefly,cloned
R4260 T6716 T6715 punct ", ",cloned
R4261 T6717 T6718 nummod 610,bp
R4262 T6718 T6715 nsubjpass bp,cloned
R4263 T6719 T6718 prep of,bp
R4264 T6720 T6721 compound mouse,cDNA
R4265 T6721 T6719 pobj cDNA,of
R4266 T6722 T6721 compound ADAM22,cDNA
R4267 T6723 T6724 punct (,1960
R4268 T6724 T6721 parataxis 1960,cDNA
R4269 T6725 T6724 dep position,1960
R4270 T6726 T6724 punct : ,1960
R4271 T6727 T6724 nummod 1351,1960
R4272 T6728 T6724 punct –,1960
R4273 T6729 T6724 prep from,1960
R4274 T6730 T6731 amod initiating,ATG
R4275 T6731 T6729 pobj ATG,from
R4276 T6732 T6724 punct ),1960
R4277 T6733 T6715 auxpass was,cloned
R4278 T6734 T6715 prep into,cloned
R4279 T6735 T6736 det the,SK
R4280 T6736 T6734 pobj SK,into
R4281 T6737 T6736 nmod pBluescript,SK
R4282 T6738 T6736 nummod II,SK
R4283 T6739 T6740 punct (,+
R4284 T6740 T6736 punct +,SK
R4285 T6741 T6740 punct ),+
R4286 T6742 T6736 cc or,SK
R4287 T6743 T6744 det the,KS
R4288 T6744 T6747 nmod KS,vector
R4289 T6745 T6744 nmod pBluescript,KS
R4290 T6746 T6744 nummod II,KS
R4291 T6747 T6736 conj vector,SK
R4292 T6748 T6749 punct (,+
R4293 T6749 T6744 punct +,KS
R4294 T6750 T6749 punct ),+
R4295 T6751 T6715 punct .,cloned
R4296 T6753 T6754 advcl Using,generated
R4297 T6755 T6756 det these,plasmids
R4298 T6756 T6753 dobj plasmids,Using
R4299 T6757 T6753 prep as,Using
R4300 T6758 T6757 pobj templates,as
R4301 T6759 T6754 punct ", ",generated
R4302 T6760 T6761 npadvmod sense,labelled
R4303 T6761 T6764 amod labelled,probes
R4304 T6762 T6760 cc and,sense
R4305 T6763 T6760 conj antisense,sense
R4306 T6764 T6754 nsubjpass probes,generated
R4307 T6765 T6764 compound RNA,probes
R4308 T6766 T6754 auxpass were,generated
R4309 T6767 T6754 agent by,generated
R4310 T6768 T6769 compound T7,polymerase
R4311 T6769 T6767 pobj polymerase,by
R4312 T6770 T6769 compound RNA,polymerase
R4313 T6771 T6769 cc and,polymerase
R4314 T6772 T6773 punct [,UTP
R4315 T6773 T6769 conj UTP,polymerase
R4316 T6774 T6773 nmod α35S,UTP
R4317 T6775 T6773 punct ],UTP
R4318 T6776 T6754 punct .,generated
R4319 T6778 T6779 amod Frozen,brain
R4320 T6779 T6780 nsubjpass brain,used
R4321 T6781 T6779 cc and,brain
R4322 T6782 T6783 amod spinal,cord
R4323 T6783 T6779 conj cord,brain
R4324 T6784 T6779 prep from,brain
R4325 T6785 T6786 nummod 2,month
R4326 T6786 T6787 npadvmod month,old
R4327 T6787 T6789 amod old,mice
R4328 T6788 T6787 punct -,old
R4329 T6789 T6784 pobj mice,from
R4330 T6790 T6789 nmod C57BL,mice
R4331 T6791 T6790 punct /,C57BL
R4332 T6792 T6790 nummod 6,C57BL
R4333 T6793 T6789 amod female,mice
R4334 T6794 T6780 auxpass were,used
R4335 T6795 T6780 prep in,used
R4336 T6796 T6797 det this,analysis
R4337 T6797 T6795 pobj analysis,in
R4338 T6798 T6780 punct .,used
R4339 T6800 T6801 nsubjpass Pretreatment,carried
R4340 T6802 T6800 punct ", ",Pretreatment
R4341 T6803 T6800 conj hybridisation,Pretreatment
R4342 T6804 T6803 punct ", ",hybridisation
R4343 T6805 T6806 compound RNase,treatment
R4344 T6806 T6803 conj treatment,hybridisation
R4345 T6807 T6806 cc and,treatment
R4346 T6808 T6806 conj washing,treatment
R4347 T6809 T6801 auxpass was,carried
R4348 T6810 T6801 prt out,carried
R4349 T6811 T6801 advcl following,carried
R4350 T6812 T6813 det the,protocol
R4351 T6813 T6811 dobj protocol,following
R4352 T6814 T6813 acl described,protocol
R4353 T6815 T6814 prep in,described
R4354 T6816 T6817 det the,literature
R4355 T6817 T6815 pobj literature,in
R4356 T6818 T6819 punct [,33
R4357 T6819 T6801 parataxis 33,carried
R4358 T6820 T6819 punct ],33
R4359 T6821 T6801 punct .,carried
R4360 T6823 T6824 amod Dehydrated,slides
R4361 T6824 T6825 nsubjpass slides,attached
R4362 T6826 T6825 auxpass were,attached
R4363 T6827 T6825 prep to,attached
R4364 T6828 T6829 compound imaging,plates
R4365 T6829 T6827 pobj plates,to
R4366 T6830 T6825 prep for,attached
R4367 T6831 T6832 nummod 48,hours
R4368 T6832 T6830 pobj hours,for
R4369 T6833 T6825 cc and,attached
R4370 T6834 T6835 det the,autoradiograms
R4371 T6835 T6836 nsubjpass autoradiograms,analysed
R4372 T6836 T6825 conj analysed,attached
R4373 T6837 T6836 auxpass were,analysed
R4374 T6838 T6836 advcl using,analysed
R4375 T6839 T6840 det a,Analyser
R4376 T6840 T6838 dobj Analyser,using
R4377 T6841 T6840 compound Bio-Image,Analyser
R4378 T6842 T6843 punct (,Film
R4379 T6843 T6840 parataxis Film,Analyser
R4380 T6844 T6843 dep BAS3000,Film
R4381 T6845 T6843 punct ", ",Film
R4382 T6846 T6843 compound Fuji,Film
R4383 T6847 T6843 compound Photo,Film
R4384 T6848 T6843 punct ),Film
R4385 T6849 T6836 punct .,analysed