0 6 Direct direct JJ 7 20 demonstration demonstration NN 21 23 of of IN 24 29 NFATp NFATp NNP 30 47 dephosphorylation dephosphorylation NN 48 51 and and CC 52 59 nuclear nuclear JJ 60 72 localization localization NN 73 75 in in IN 76 85 activated activate VBN 86 90 HT-2 ht-2 NN 91 96 cells cell NNS 97 102 using use VBG 103 104 a a DT 105 113 specific specific JJ 114 119 NFATp NFATp NNP 120 130 polyclonal polyclonal JJ 131 139 antibody antibody NN 139 140 . . . 142 149 Nuclear nuclear JJ 150 156 factor factor NN 157 159 of of IN 160 169 activated activate VBN 170 171 T t NN 172 177 cells cell NNS 178 179 ( ( ( 179 183 NFAT NFAT NNP 183 184 ) ) ) 185 194 regulates regulate VBZ 195 208 transcription transcription NN 209 211 of of IN 212 213 a a DT 214 220 number number NN 221 223 of of IN 224 232 cytokine cytokine NN 233 238 genes gene NNS 238 239 , , , 240 243 and and CC 244 248 NFAT NFAT NNP 249 252 DNA DNA NNP 253 260 binding binding NN 261 269 activity activity NN 270 272 is be VBZ 273 283 stimulated stimulate VBN 284 293 following follow VBG 294 295 T t NN 296 300 cell cell NN 301 311 activation activation NN 311 312 . . . 313 320 Several several JJ 321 326 lines line NNS 327 329 of of IN 330 338 evidence evidence NN 339 343 have have VBP 344 353 suggested suggest VBN 354 358 that that IN 359 363 NFAT NFAT NNP 364 366 is be VBZ 367 368 a a DT 369 378 substrate substrate NN 379 382 for for IN 383 394 calcineurin calcineurin NN 394 395 , , , 396 397 a a DT 398 414 serine/threonine serine/threonine NN 415 426 phosphatase phosphatase NN 426 427 . . . 428 433 Using use VBG 434 435 a a DT 436 446 polyclonal polyclonal JJ 447 455 antibody antibody NN 456 458 to to TO 459 465 murine murine JJ 466 471 NFATp NFATp NNP 471 472 , , , 473 480 Western western NN 481 485 blot blot NN 486 494 analysis analysis NN 495 497 of of IN 498 505 various various JJ 506 511 mouse mouse NN 512 519 tissues tissue NNS 520 532 demonstrated demonstrate VBD 533 537 that that IN 538 541 the the DT 542 553 110-130-kDa 110-130-kda JJ 554 559 NFATp NFATp NNP 560 567 protein protein NN 568 571 was be VBD 572 578 highly highly RB 579 588 expressed express VBN 589 591 in in IN 592 598 thymus thymus NN 599 602 and and CC 603 609 spleen spleen NN 609 610 . . . 611 620 Treatment Treatment NNP 621 623 of of IN 624 642 immunoprecipitated immunoprecipitated JJ 643 648 NFATp NFATp NNP 649 653 from from IN 654 663 untreated untreated JJ 664 668 HT-2 ht-2 NN 669 674 cells cell NNS 675 679 with with IN 680 691 calcineurin calcineurin NN 692 700 resulted result VBD 701 703 in in IN 704 707 the the DT 708 725 dephosphorylation dephosphorylation NN 726 728 of of IN 729 734 NFATp NFATp NNP 734 735 , , , 736 749 demonstrating demonstrate VBG 750 754 that that IN 755 760 NFATp NFATp NNP 761 763 is be VBZ 764 766 an an DT 767 769 in in FW 770 775 vitro vitro FW 776 785 substrate substrate NN 786 789 for for IN 790 801 calcineurin calcineurin NN 801 802 . . . 803 808 NFATp NFATp NNP 809 827 immunoprecipitated immunoprecipitate VBN 828 832 from from IN 833 844 32P-labeled 32p-labeled JJ 845 849 HT-2 ht-2 NN 850 855 cells cell NNS 856 864 migrated migrate VBD 865 867 as as IN 868 870 an an DT 871 884 approximately approximately RB 885 892 120-kDa 120-kda JJ 893 900 protein protein NN 901 905 that that WDT 906 909 was be VBD 910 919 localized localized JJ 920 922 to to TO 923 926 the the DT 927 934 cytosol cytosol NN 935 937 of of IN 938 941 the the DT 942 947 cells cell NNS 947 948 . . . 949 958 Treatment Treatment NNP 959 961 of of IN 962 965 the the DT 966 971 cells cell NNS 972 976 with with IN 977 986 ionomycin ionomycin NN 987 995 resulted result VBD 996 998 in in IN 999 1000 a a DT 1001 1009 decrease decrease NN 1010 1012 in in IN 1013 1016 the the DT 1017 1026 molecular molecular JJ 1027 1033 weight weight NN 1034 1036 of of IN 1037 1042 NFATp NFATp NNP 1043 1046 and and CC 1047 1048 a a DT 1049 1053 loss loss NN 1054 1056 of of IN 1057 1060 32P 32p NN 1060 1061 , , , 1062 1072 consistent consistent JJ 1073 1077 with with IN 1078 1083 NFATp NFATp NNP 1084 1101 dephosphorylation dephosphorylation NN 1101 1102 . . . 1103 1106 The the DT 1107 1124 dephosphorylation dephosphorylation NN 1125 1127 of of IN 1128 1133 NFATp NFATp NNP 1134 1137 was be VBD 1138 1149 accompanied accompany VBN 1150 1152 by by IN 1153 1165 localization localization NN 1166 1168 of of IN 1169 1172 the the DT 1173 1180 protein protein NN 1181 1183 to to TO 1184 1187 the the DT 1188 1195 nuclear nuclear JJ 1196 1204 fraction fraction NN 1204 1205 . . . 1206 1210 Both both DT 1211 1213 of of IN 1214 1219 these these DT 1220 1226 events event NNS 1227 1231 were be VBD 1232 1239 blocked block VBN 1240 1242 by by IN 1243 1256 preincubation preincubation NN 1257 1259 of of IN 1260 1263 the the DT 1264 1269 cells cell NNS 1270 1274 with with IN 1275 1280 FK506 FK506 NNP 1280 1281 , , , 1282 1283 a a DT 1284 1295 calcineurin calcineurin NN 1296 1305 inhibitor inhibitor NN 1305 1306 , , , 1307 1317 consistent consistent JJ 1318 1322 with with IN 1323 1326 the the DT 1327 1337 hypothesis hypothesis NN 1338 1342 that that IN 1343 1348 NFATp NFATp NNP 1349 1351 is be VBZ 1352 1353 a a DT 1354 1365 calcineurin calcineurin NN 1366 1375 substrate substrate NN 1376 1378 in in IN 1379 1384 cells cell NNS 1384 1385 . . .