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PMC:1087847 / 36570-37999 JSONTXT 5 Projects

Annnotations TAB TSV DIC JSON TextAE

Id Subject Object Predicate Lexical cue
T10539 0-10 NN denotes Extraction
T10540 11-13 IN denotes of
T10541 14-21 JJ denotes nuclear
T10543 22-29 NN denotes Annexin
T10542 30-32 NN denotes A7
T10544 32-97 sentence denotes Cultured cells were trypsinized, counted and washed once in PBS.
T10545 33-41 VBN denotes Cultured
T10546 42-47 NNS denotes cells
T10548 48-52 VBD denotes were
T10547 53-64 VBN denotes trypsinized
T10549 64-66 , denotes ,
T10550 66-73 VBN denotes counted
T10551 74-77 CC denotes and
T10552 78-84 VBN denotes washed
T10553 85-89 RB denotes once
T10554 90-92 IN denotes in
T10555 93-96 NN denotes PBS
T10556 96-97 . denotes .
T10557 97-354 sentence denotes They were resuspended in buffer1 (0.32 M sucrose, 10 mM Tris, pH8.0, 3 mM CaCl2, 2 mM Mg-acetate, 0.1 mM EDTA, 0.5% NP40, 1 mM DTT, 0.5 mM PMSF; 1 × 107cells/400μl buffer) and sonified on ice (sonifier UP200S, dr.hielscher, 40 s, amplitude 50%, cycle 0.5).
T10558 98-102 PRP denotes They
T10560 103-107 VBD denotes were
T10559 108-119 VBN denotes resuspended
T10561 120-122 IN denotes in
T10562 123-130 NN denotes buffer1
T10563 131-132 -LRB- denotes (
T10565 132-136 CD denotes 0.32
T10566 137-138 NN denotes M
T10564 139-146 NN denotes sucrose
T10567 146-148 , denotes ,
T10568 148-150 CD denotes 10
T10569 151-153 NN denotes mM
T10570 154-158 NN denotes Tris
T10571 158-160 , denotes ,
T10572 160-165 NN denotes pH8.0
T10573 165-167 , denotes ,
T10574 167-168 CD denotes 3
T10575 169-171 NN denotes mM
T10576 172-177 NN denotes CaCl2
T10577 177-179 , denotes ,
T10578 179-180 CD denotes 2
T10579 181-183 NN denotes mM
T10581 184-186 NN denotes Mg
T10582 186-187 HYPH denotes -
T10580 187-194 NN denotes acetate
T10583 194-196 , denotes ,
T10584 196-199 CD denotes 0.1
T10585 200-202 NN denotes mM
T10586 203-207 NN denotes EDTA
T10587 207-209 , denotes ,
T10588 209-212 CD denotes 0.5
T10589 212-213 NN denotes %
T10590 214-218 NN denotes NP40
T10591 218-220 , denotes ,
T10592 220-221 CD denotes 1
T10593 222-224 NN denotes mM
T10594 225-228 NN denotes DTT
T10595 228-230 , denotes ,
T10596 230-233 CD denotes 0.5
T10597 234-236 NN denotes mM
T10598 237-241 NN denotes PMSF
T10599 241-242 : denotes ;
T10600 243-244 CD denotes 1
T10602 245-246 SYM denotes ×
T10601 247-255 NNS denotes 107cells
T10603 255-256 SYM denotes /
T10605 256-261 NN denotes 400μl
T10604 262-268 NN denotes buffer
T10606 268-269 -RRB- denotes )
T10607 270-273 CC denotes and
T10608 274-282 VBN denotes sonified
T10609 283-285 IN denotes on
T10610 286-289 NN denotes ice
T10611 290-291 -LRB- denotes (
T10613 291-299 NN denotes sonifier
T10614 300-306 NN denotes UP200S
T10615 306-308 , denotes ,
T10616 308-320 NNP denotes dr.hielscher
T10617 320-322 , denotes ,
T10618 322-324 CD denotes 40
T10619 325-326 NN denotes s
T10620 326-328 , denotes ,
T10621 328-337 NN denotes amplitude
T10623 338-340 CD denotes 50
T10622 340-341 NN denotes %
T10624 341-343 , denotes ,
T10612 343-348 NN denotes cycle
T10625 349-352 CD denotes 0.5
T10626 352-353 -RRB- denotes )
T10627 353-354 . denotes .
T10628 354-439 sentence denotes This step was repeated until no more intact cells were observed by light microscopy.
T10629 355-359 DT denotes This
T10630 360-364 NN denotes step
T10632 365-368 VBD denotes was
T10631 369-377 VBN denotes repeated
T10633 378-383 IN denotes until
T10635 384-386 DT denotes no
T10637 387-391 JJR denotes more
T10638 392-398 JJ denotes intact
T10636 399-404 NNS denotes cells
T10639 405-409 VBD denotes were
T10634 410-418 VBN denotes observed
T10640 419-421 IN denotes by
T10641 422-427 NN denotes light
T10642 428-438 NN denotes microscopy
T10643 438-439 . denotes .
T10644 439-570 sentence denotes The resulting homogenate was centrifuged at 500 g for 5 minutes, washed twice in 1 ml buffer 1 without NP40 and centrifuged again.
T10645 440-443 DT denotes The
T10647 444-453 VBG denotes resulting
T10646 454-464 NN denotes homogenate
T10649 465-468 VBD denotes was
T10648 469-480 VBN denotes centrifuged
T10650 481-483 IN denotes at
T10651 484-487 CD denotes 500
T10652 488-489 NN denotes g
T10653 490-493 IN denotes for
T10654 494-495 CD denotes 5
T10655 496-503 NNS denotes minutes
T10656 503-505 , denotes ,
T10657 505-511 VBN denotes washed
T10658 512-517 RB denotes twice
T10659 518-520 IN denotes in
T10660 521-522 CD denotes 1
T10661 523-525 NN denotes ml
T10662 526-532 NN denotes buffer
T10663 533-534 CD denotes 1
T10664 535-542 IN denotes without
T10665 543-547 NN denotes NP40
T10666 548-551 CC denotes and
T10667 552-563 VBN denotes centrifuged
T10668 564-569 RB denotes again
T10669 569-570 . denotes .
T10670 570-617 sentence denotes The resulting pellet contains purified nuclei.
T10671 571-574 DT denotes The
T10673 575-584 VBG denotes resulting
T10672 585-591 NN denotes pellet
T10674 592-600 VBZ denotes contains
T10675 601-609 VBN denotes purified
T10676 610-616 NNS denotes nuclei
T10677 616-617 . denotes .
T10678 617-887 sentence denotes For partial extraction of the nucleoplasm according to [37], nuclei were resuspended in buffer 2 (20 mM Hepes, pH 7.9, 1.5 mM MgCl2, 20 mM KCl, 0.2 mM EDTA, 25%v/v glycerine, 0.5 mM DTT, 0.5 mM PMSF; nuclei of 107cells/30 μl buffer) and incubated on ice for 30 minutes.
T10679 618-621 IN denotes For
T10681 622-629 JJ denotes partial
T10682 630-640 NN denotes extraction
T10683 641-643 IN denotes of
T10684 644-647 DT denotes the
T10685 648-659 NN denotes nucleoplasm
T10686 660-669 VBG denotes according
T10687 670-672 IN denotes to
T10688 673-674 -LRB- denotes [
T10689 674-676 CD denotes 37
T10690 676-677 -RRB- denotes ]
T10691 677-679 , denotes ,
T10692 679-685 NNS denotes nuclei
T10693 686-690 VBD denotes were
T10680 691-702 VBN denotes resuspended
T10694 703-705 IN denotes in
T10695 706-712 NN denotes buffer
T10696 713-714 CD denotes 2
T10697 715-716 -LRB- denotes (
T10699 716-718 CD denotes 20
T10700 719-721 NN denotes mM
T10698 722-727 NN denotes Hepes
T10701 727-729 , denotes ,
T10702 729-731 NN denotes pH
T10703 732-735 CD denotes 7.9
T10704 735-737 , denotes ,
T10705 737-740 CD denotes 1.5
T10706 741-743 NN denotes mM
T10707 744-749 NN denotes MgCl2
T10708 749-751 , denotes ,
T10709 751-753 CD denotes 20
T10710 754-756 NN denotes mM
T10711 757-760 NN denotes KCl
T10712 760-762 , denotes ,
T10713 762-765 CD denotes 0.2
T10714 766-768 NN denotes mM
T10715 769-773 NN denotes EDTA
T10716 773-775 , denotes ,
T10717 775-777 CD denotes 25
T10718 777-778 NN denotes %
T10720 778-779 NN denotes v
T10721 779-780 SYM denotes /
T10722 780-781 NN denotes v
T10719 782-791 NN denotes glycerine
T10723 791-793 , denotes ,
T10724 793-796 CD denotes 0.5
T10725 797-799 NN denotes mM
T10726 800-803 NN denotes DTT
T10727 803-805 , denotes ,
T10728 805-808 CD denotes 0.5
T10729 809-811 NN denotes mM
T10730 812-816 NN denotes PMSF
T10731 816-817 : denotes ;
T10732 818-824 NNS denotes nuclei
T10733 825-827 IN denotes of
T10734 828-836 NNS denotes 107cells
T10736 836-837 SYM denotes /
T10738 837-839 CD denotes 30
T10737 840-842 NN denotes μl
T10735 843-849 NN denotes buffer
T10739 849-850 -RRB- denotes )
T10740 851-854 CC denotes and
T10741 855-864 VBN denotes incubated
T10742 865-867 IN denotes on
T10743 868-871 NN denotes ice
T10744 872-875 IN denotes for
T10745 876-878 CD denotes 30
T10746 879-886 NNS denotes minutes
T10747 886-887 . denotes .
T10748 887-1163 sentence denotes An equal volume of buffer 3 was added in aliquots (20 mM Hepes, pH 7.9, 1.5 mM MgCl2, 800 mM KCl, 0.2 mM EDTA, 25%v/v glycerine, 0.5 mM DTT, 0.5 mM PMSF, 1% NP40, protease inhibitor cocktail (Sigma)) and the solution was incubated while shaking for further 30 minutes on ice.
T10749 888-890 DT denotes An
T10751 891-896 JJ denotes equal
T10750 897-903 NN denotes volume
T10753 904-906 IN denotes of
T10754 907-913 NN denotes buffer
T10755 914-915 CD denotes 3
T10756 916-919 VBD denotes was
T10752 920-925 VBN denotes added
T10757 926-928 IN denotes in
T10758 929-937 NNS denotes aliquots
T10759 938-939 -LRB- denotes (
T10761 939-941 CD denotes 20
T10762 942-944 NN denotes mM
T10763 945-950 NN denotes Hepes
T10764 950-952 , denotes ,
T10765 952-954 NN denotes pH
T10766 955-958 CD denotes 7.9
T10767 958-960 , denotes ,
T10768 960-963 CD denotes 1.5
T10769 964-966 NN denotes mM
T10770 967-972 NN denotes MgCl2
T10771 972-974 , denotes ,
T10772 974-977 CD denotes 800
T10773 978-980 NN denotes mM
T10774 981-984 NN denotes KCl
T10775 984-986 , denotes ,
T10776 986-989 CD denotes 0.2
T10777 990-992 NN denotes mM
T10778 993-997 NN denotes EDTA
T10779 997-999 , denotes ,
T10780 999-1001 CD denotes 25
T10781 1001-1002 NN denotes %
T10783 1002-1003 NN denotes v
T10784 1003-1004 SYM denotes /
T10785 1004-1005 NN denotes v
T10782 1006-1015 NN denotes glycerine
T10786 1015-1017 , denotes ,
T10787 1017-1020 CD denotes 0.5
T10788 1021-1023 NN denotes mM
T10789 1024-1027 NN denotes DTT
T10790 1027-1029 , denotes ,
T10791 1029-1032 CD denotes 0.5
T10792 1033-1035 NN denotes mM
T10793 1036-1040 NN denotes PMSF
T10794 1040-1042 , denotes ,
T10795 1042-1043 CD denotes 1
T10796 1043-1044 NN denotes %
T10797 1045-1049 NN denotes NP40
T10798 1049-1051 , denotes ,
T10799 1051-1059 NN denotes protease
T10800 1060-1069 NN denotes inhibitor
T10801 1070-1078 NN denotes cocktail
T10802 1079-1080 -LRB- denotes (
T10760 1080-1085 NNP denotes Sigma
T10803 1085-1086 -RRB- denotes )
T10804 1086-1087 -RRB- denotes )
T10805 1088-1091 CC denotes and
T10806 1092-1095 DT denotes the
T10807 1096-1104 NN denotes solution
T10809 1105-1108 VBD denotes was
T10808 1109-1118 VBN denotes incubated
T10810 1119-1124 IN denotes while
T10811 1125-1132 VBG denotes shaking
T10812 1133-1136 IN denotes for
T10813 1137-1144 JJ denotes further
T10815 1145-1147 CD denotes 30
T10814 1148-1155 NNS denotes minutes
T10816 1156-1158 IN denotes on
T10817 1159-1162 NN denotes ice
T10818 1162-1163 . denotes .
T10819 1163-1349 sentence denotes Then the solution was centrifuged for 15 minutes at 14.000 g, the resulting supernatant was used as nucleoplasm, and the pellet was washed once and adjusted to an equal volume with PBS.
T10820 1164-1168 RB denotes Then
T10822 1169-1172 DT denotes the
T10823 1173-1181 NN denotes solution
T10824 1182-1185 VBD denotes was
T10821 1186-1197 VBN denotes centrifuged
T10825 1198-1201 IN denotes for
T10826 1202-1204 CD denotes 15
T10827 1205-1212 NNS denotes minutes
T10828 1213-1215 IN denotes at
T10829 1216-1222 CD denotes 14.000
T10830 1223-1224 NN denotes g
T10831 1224-1226 , denotes ,
T10832 1226-1229 DT denotes the
T10834 1230-1239 VBG denotes resulting
T10833 1240-1251 NN denotes supernatant
T10836 1252-1255 VBD denotes was
T10835 1256-1260 VBN denotes used
T10837 1261-1263 IN denotes as
T10838 1264-1275 NN denotes nucleoplasm
T10839 1275-1277 , denotes ,
T10840 1277-1280 CC denotes and
T10841 1281-1284 DT denotes the
T10842 1285-1291 NN denotes pellet
T10844 1292-1295 VBD denotes was
T10843 1296-1302 VBN denotes washed
T10845 1303-1307 RB denotes once
T10846 1308-1311 CC denotes and
T10847 1312-1320 VBN denotes adjusted
T10848 1321-1323 IN denotes to
T10849 1324-1326 DT denotes an
T10851 1327-1332 JJ denotes equal
T10850 1333-1339 NN denotes volume
T10852 1340-1344 IN denotes with
T10853 1345-1348 NN denotes PBS
T10854 1348-1349 . denotes .
T10855 1349-1429 sentence denotes Nucleoplasm and nuclear pellet were subjected to SDS-PAGE and Western blotting.
T10856 1350-1361 NN denotes Nucleoplasm
T10858 1362-1365 CC denotes and
T10859 1366-1373 JJ denotes nuclear
T10860 1374-1380 NN denotes pellet
T10861 1381-1385 VBD denotes were
T10857 1386-1395 VBN denotes subjected
T10862 1396-1398 IN denotes to
T10863 1399-1402 NN denotes SDS
T10865 1402-1403 HYPH denotes -
T10864 1403-1407 NN denotes PAGE
T10866 1408-1411 CC denotes and
T10867 1412-1419 NNP denotes Western
T10868 1420-1428 NN denotes blotting
T10869 1428-1429 . denotes .
R5065 T10540 T10539 prep of,Extraction
R5066 T10541 T10542 amod nuclear,A7
R5067 T10542 T10540 pobj A7,of
R5068 T10543 T10542 compound Annexin,A7
R5069 T10545 T10546 amod Cultured,cells
R5070 T10546 T10547 nsubjpass cells,trypsinized
R5071 T10548 T10547 auxpass were,trypsinized
R5072 T10549 T10547 punct ", ",trypsinized
R5073 T10550 T10547 conj counted,trypsinized
R5074 T10551 T10550 cc and,counted
R5075 T10552 T10550 conj washed,counted
R5076 T10553 T10552 advmod once,washed
R5077 T10554 T10552 prep in,washed
R5078 T10555 T10554 pobj PBS,in
R5079 T10556 T10547 punct .,trypsinized
R5080 T10558 T10559 nsubjpass They,resuspended
R5081 T10560 T10559 auxpass were,resuspended
R5082 T10561 T10559 prep in,resuspended
R5083 T10562 T10561 pobj buffer1,in
R5084 T10563 T10564 punct (,sucrose
R5085 T10564 T10562 parataxis sucrose,buffer1
R5086 T10565 T10566 nummod 0.32,M
R5087 T10566 T10564 compound M,sucrose
R5088 T10567 T10564 punct ", ",sucrose
R5089 T10568 T10569 nummod 10,mM
R5090 T10569 T10570 compound mM,Tris
R5091 T10570 T10564 appos Tris,sucrose
R5092 T10571 T10570 punct ", ",Tris
R5093 T10572 T10570 npadvmod pH8.0,Tris
R5094 T10573 T10564 punct ", ",sucrose
R5095 T10574 T10575 nummod 3,mM
R5096 T10575 T10576 compound mM,CaCl2
R5097 T10576 T10564 appos CaCl2,sucrose
R5098 T10577 T10564 punct ", ",sucrose
R5099 T10578 T10579 nummod 2,mM
R5100 T10579 T10580 compound mM,acetate
R5101 T10580 T10564 appos acetate,sucrose
R5102 T10581 T10580 compound Mg,acetate
R5103 T10582 T10580 punct -,acetate
R5104 T10583 T10564 punct ", ",sucrose
R5105 T10584 T10585 nummod 0.1,mM
R5106 T10585 T10586 compound mM,EDTA
R5107 T10586 T10564 appos EDTA,sucrose
R5108 T10587 T10564 punct ", ",sucrose
R5109 T10588 T10589 nummod 0.5,%
R5110 T10589 T10590 compound %,NP40
R5111 T10590 T10564 appos NP40,sucrose
R5112 T10591 T10564 punct ", ",sucrose
R5113 T10592 T10593 nummod 1,mM
R5114 T10593 T10594 compound mM,DTT
R5115 T10594 T10564 appos DTT,sucrose
R5116 T10595 T10564 punct ", ",sucrose
R5117 T10596 T10597 nummod 0.5,mM
R5118 T10597 T10598 compound mM,PMSF
R5119 T10598 T10564 appos PMSF,sucrose
R5120 T10599 T10564 punct ;,sucrose
R5121 T10600 T10601 nummod 1,107cells
R5122 T10601 T10564 appos 107cells,sucrose
R5123 T10602 T10600 punct ×,1
R5124 T10603 T10604 punct /,buffer
R5125 T10604 T10601 prep buffer,107cells
R5126 T10605 T10604 compound 400μl,buffer
R5127 T10606 T10564 punct ),sucrose
R5128 T10607 T10559 cc and,resuspended
R5129 T10608 T10559 conj sonified,resuspended
R5130 T10609 T10608 prep on,sonified
R5131 T10610 T10609 pobj ice,on
R5132 T10611 T10612 punct (,cycle
R5133 T10612 T10608 parataxis cycle,sonified
R5134 T10613 T10614 compound sonifier,UP200S
R5135 T10614 T10612 dep UP200S,cycle
R5136 T10615 T10612 punct ", ",cycle
R5137 T10616 T10612 dep dr.hielscher,cycle
R5138 T10617 T10612 punct ", ",cycle
R5139 T10618 T10619 nummod 40,s
R5140 T10619 T10612 dep s,cycle
R5141 T10620 T10612 punct ", ",cycle
R5142 T10621 T10622 dep amplitude,%
R5143 T10622 T10612 dep %,cycle
R5144 T10623 T10622 nummod 50,%
R5145 T10624 T10612 punct ", ",cycle
R5146 T10625 T10612 nummod 0.5,cycle
R5147 T10626 T10612 punct ),cycle
R5148 T10627 T10559 punct .,resuspended
R5149 T10629 T10630 det This,step
R5150 T10630 T10631 nsubjpass step,repeated
R5151 T10632 T10631 auxpass was,repeated
R5152 T10633 T10634 mark until,observed
R5153 T10634 T10631 advcl observed,repeated
R5154 T10635 T10636 det no,cells
R5155 T10636 T10634 nsubjpass cells,observed
R5156 T10637 T10636 amod more,cells
R5157 T10638 T10636 amod intact,cells
R5158 T10639 T10634 auxpass were,observed
R5159 T10640 T10634 prep by,observed
R5160 T10641 T10642 compound light,microscopy
R5161 T10642 T10640 pobj microscopy,by
R5162 T10643 T10631 punct .,repeated
R5163 T10645 T10646 det The,homogenate
R5164 T10646 T10648 nsubjpass homogenate,centrifuged
R5165 T10647 T10646 amod resulting,homogenate
R5166 T10649 T10648 auxpass was,centrifuged
R5167 T10650 T10648 prep at,centrifuged
R5168 T10651 T10652 nummod 500,g
R5169 T10652 T10650 pobj g,at
R5170 T10653 T10648 prep for,centrifuged
R5171 T10654 T10655 nummod 5,minutes
R5172 T10655 T10653 pobj minutes,for
R5173 T10656 T10648 punct ", ",centrifuged
R5174 T10657 T10648 conj washed,centrifuged
R5175 T10658 T10657 advmod twice,washed
R5176 T10659 T10657 prep in,washed
R5177 T10660 T10661 nummod 1,ml
R5178 T10661 T10662 compound ml,buffer
R5179 T10662 T10659 pobj buffer,in
R5180 T10663 T10662 nummod 1,buffer
R5181 T10664 T10657 prep without,washed
R5182 T10665 T10664 pobj NP40,without
R5183 T10666 T10657 cc and,washed
R5184 T10667 T10657 conj centrifuged,washed
R5185 T10668 T10667 advmod again,centrifuged
R5186 T10669 T10648 punct .,centrifuged
R5187 T10671 T10672 det The,pellet
R5188 T10672 T10674 nsubj pellet,contains
R5189 T10673 T10672 amod resulting,pellet
R5190 T10675 T10676 amod purified,nuclei
R5191 T10676 T10674 dobj nuclei,contains
R5192 T10677 T10674 punct .,contains
R5193 T10679 T10680 prep For,resuspended
R5194 T10681 T10682 amod partial,extraction
R5195 T10682 T10679 pobj extraction,For
R5196 T10683 T10682 prep of,extraction
R5197 T10684 T10685 det the,nucleoplasm
R5198 T10685 T10683 pobj nucleoplasm,of
R5199 T10686 T10682 prep according,extraction
R5200 T10687 T10686 prep to,according
R5201 T10688 T10687 punct [,to
R5202 T10689 T10687 pobj 37,to
R5203 T10690 T10680 punct ],resuspended
R5204 T10691 T10680 punct ", ",resuspended
R5205 T10692 T10680 nsubjpass nuclei,resuspended
R5206 T10693 T10680 auxpass were,resuspended
R5207 T10694 T10680 prep in,resuspended
R5208 T10695 T10694 pobj buffer,in
R5209 T10696 T10695 nummod 2,buffer
R5210 T10697 T10698 punct (,Hepes
R5211 T10698 T10695 parataxis Hepes,buffer
R5212 T10699 T10700 nummod 20,mM
R5213 T10700 T10698 compound mM,Hepes
R5214 T10701 T10698 punct ", ",Hepes
R5215 T10702 T10698 npadvmod pH,Hepes
R5216 T10703 T10702 nummod 7.9,pH
R5217 T10704 T10698 punct ", ",Hepes
R5218 T10705 T10706 nummod 1.5,mM
R5219 T10706 T10707 compound mM,MgCl2
R5220 T10707 T10698 appos MgCl2,Hepes
R5221 T10708 T10698 punct ", ",Hepes
R5222 T10709 T10710 nummod 20,mM
R5223 T10710 T10711 compound mM,KCl
R5224 T10711 T10698 appos KCl,Hepes
R5225 T10712 T10698 punct ", ",Hepes
R5226 T10713 T10714 nummod 0.2,mM
R5227 T10714 T10715 compound mM,EDTA
R5228 T10715 T10698 appos EDTA,Hepes
R5229 T10716 T10698 punct ", ",Hepes
R5230 T10717 T10718 nummod 25,%
R5231 T10718 T10719 nmod %,glycerine
R5232 T10719 T10698 appos glycerine,Hepes
R5233 T10720 T10718 appos v,%
R5234 T10721 T10722 punct /,v
R5235 T10722 T10720 prep v,v
R5236 T10723 T10698 punct ", ",Hepes
R5237 T10724 T10725 nummod 0.5,mM
R5238 T10725 T10726 compound mM,DTT
R5239 T10726 T10698 appos DTT,Hepes
R5240 T10727 T10698 punct ", ",Hepes
R5241 T10728 T10729 nummod 0.5,mM
R5242 T10729 T10730 compound mM,PMSF
R5243 T10730 T10698 appos PMSF,Hepes
R5244 T10731 T10698 punct ;,Hepes
R5245 T10732 T10698 appos nuclei,Hepes
R5246 T10733 T10732 prep of,nuclei
R5247 T10734 T10735 nmod 107cells,buffer
R5248 T10735 T10733 pobj buffer,of
R5249 T10736 T10737 punct /,μl
R5250 T10737 T10734 prep μl,107cells
R5251 T10738 T10737 nummod 30,μl
R5252 T10739 T10698 punct ),Hepes
R5253 T10740 T10680 cc and,resuspended
R5254 T10741 T10680 conj incubated,resuspended
R5255 T10742 T10741 prep on,incubated
R5256 T10743 T10742 pobj ice,on
R5257 T10744 T10741 prep for,incubated
R5258 T10745 T10746 nummod 30,minutes
R5259 T10746 T10744 pobj minutes,for
R5260 T10747 T10680 punct .,resuspended
R5261 T10749 T10750 det An,volume
R5262 T10750 T10752 nsubjpass volume,added
R5263 T10751 T10750 amod equal,volume
R5264 T10753 T10750 prep of,volume
R5265 T10754 T10753 pobj buffer,of
R5266 T10755 T10754 nummod 3,buffer
R5267 T10756 T10752 auxpass was,added
R5268 T10757 T10752 prep in,added
R5269 T10758 T10757 pobj aliquots,in
R5270 T10759 T10760 punct (,Sigma
R5271 T10760 T10752 parataxis Sigma,added
R5272 T10761 T10762 nummod 20,mM
R5273 T10762 T10763 compound mM,Hepes
R5274 T10763 T10760 dep Hepes,Sigma
R5275 T10764 T10763 punct ", ",Hepes
R5276 T10765 T10763 npadvmod pH,Hepes
R5277 T10766 T10765 nummod 7.9,pH
R5278 T10767 T10763 punct ", ",Hepes
R5279 T10768 T10769 nummod 1.5,mM
R5280 T10769 T10770 compound mM,MgCl2
R5281 T10770 T10763 appos MgCl2,Hepes
R5282 T10771 T10763 punct ", ",Hepes
R5283 T10772 T10773 nummod 800,mM
R5284 T10773 T10774 compound mM,KCl
R5285 T10774 T10763 appos KCl,Hepes
R5286 T10775 T10763 punct ", ",Hepes
R5287 T10776 T10777 nummod 0.2,mM
R5288 T10777 T10778 compound mM,EDTA
R5289 T10778 T10763 appos EDTA,Hepes
R5290 T10779 T10763 punct ", ",Hepes
R5291 T10780 T10781 nummod 25,%
R5292 T10781 T10782 nmod %,glycerine
R5293 T10782 T10763 appos glycerine,Hepes
R5294 T10783 T10781 appos v,%
R5295 T10784 T10785 punct /,v
R5296 T10785 T10783 prep v,v
R5297 T10786 T10763 punct ", ",Hepes
R5298 T10787 T10788 nummod 0.5,mM
R5299 T10788 T10789 compound mM,DTT
R5300 T10789 T10763 appos DTT,Hepes
R5301 T10790 T10763 punct ", ",Hepes
R5302 T10791 T10792 nummod 0.5,mM
R5303 T10792 T10793 compound mM,PMSF
R5304 T10793 T10763 appos PMSF,Hepes
R5305 T10794 T10763 punct ", ",Hepes
R5306 T10795 T10796 nummod 1,%
R5307 T10796 T10797 compound %,NP40
R5308 T10797 T10763 appos NP40,Hepes
R5309 T10798 T10763 punct ", ",Hepes
R5310 T10799 T10800 compound protease,inhibitor
R5311 T10800 T10801 compound inhibitor,cocktail
R5312 T10801 T10763 appos cocktail,Hepes
R5313 T10802 T10760 punct (,Sigma
R5314 T10803 T10760 punct ),Sigma
R5315 T10804 T10760 punct ),Sigma
R5316 T10805 T10752 cc and,added
R5317 T10806 T10807 det the,solution
R5318 T10807 T10808 nsubjpass solution,incubated
R5319 T10808 T10752 conj incubated,added
R5320 T10809 T10808 auxpass was,incubated
R5321 T10810 T10811 mark while,shaking
R5322 T10811 T10808 advcl shaking,incubated
R5323 T10812 T10811 prep for,shaking
R5324 T10813 T10814 amod further,minutes
R5325 T10814 T10812 pobj minutes,for
R5326 T10815 T10814 nummod 30,minutes
R5327 T10816 T10811 prep on,shaking
R5328 T10817 T10816 pobj ice,on
R5329 T10818 T10808 punct .,incubated
R5330 T10820 T10821 advmod Then,centrifuged
R5331 T10822 T10823 det the,solution
R5332 T10823 T10821 nsubjpass solution,centrifuged
R5333 T10824 T10821 auxpass was,centrifuged
R5334 T10825 T10821 prep for,centrifuged
R5335 T10826 T10827 nummod 15,minutes
R5336 T10827 T10825 pobj minutes,for
R5337 T10828 T10821 prep at,centrifuged
R5338 T10829 T10830 nummod 14.000,g
R5339 T10830 T10828 pobj g,at
R5340 T10831 T10821 punct ", ",centrifuged
R5341 T10832 T10833 det the,supernatant
R5342 T10833 T10835 nsubjpass supernatant,used
R5343 T10834 T10833 amod resulting,supernatant
R5344 T10835 T10821 conj used,centrifuged
R5345 T10836 T10835 auxpass was,used
R5346 T10837 T10835 prep as,used
R5347 T10838 T10837 pobj nucleoplasm,as
R5348 T10839 T10835 punct ", ",used
R5349 T10840 T10835 cc and,used
R5350 T10841 T10842 det the,pellet
R5351 T10842 T10843 nsubjpass pellet,washed
R5352 T10843 T10835 conj washed,used
R5353 T10844 T10843 auxpass was,washed
R5354 T10845 T10843 advmod once,washed
R5355 T10846 T10843 cc and,washed
R5356 T10847 T10843 conj adjusted,washed
R5357 T10848 T10847 prep to,adjusted
R5358 T10849 T10850 det an,volume
R5359 T10850 T10848 pobj volume,to
R5360 T10851 T10850 amod equal,volume
R5361 T10852 T10847 prep with,adjusted
R5362 T10853 T10852 pobj PBS,with
R5363 T10854 T10843 punct .,washed
R5364 T10856 T10857 nsubjpass Nucleoplasm,subjected
R5365 T10858 T10856 cc and,Nucleoplasm
R5366 T10859 T10860 amod nuclear,pellet
R5367 T10860 T10856 conj pellet,Nucleoplasm
R5368 T10861 T10857 auxpass were,subjected
R5369 T10862 T10857 prep to,subjected
R5370 T10863 T10864 compound SDS,PAGE
R5371 T10864 T10862 pobj PAGE,to
R5372 T10865 T10864 punct -,PAGE
R5373 T10866 T10864 cc and,PAGE
R5374 T10867 T10868 compound Western,blotting
R5375 T10868 T10864 conj blotting,PAGE
R5376 T10869 T10857 punct .,subjected