0 10 Functional functional JJ 11 27 characterization characterization NN 28 30 of of IN 31 36 novel novel JJ 37 41 IL-2 il-2 NN 42 57 transcriptional transcriptional JJ 58 68 inhibitors inhibitor NNS 68 69 . . . 71 84 IL-2-mediated il-2-mediated JJ 85 86 T t NN 87 91 cell cell NN 92 105 proliferation proliferation NN 106 108 is be VBZ 109 110 a a DT 111 119 critical critical JJ 120 125 early early JJ 126 131 event event NN 132 134 in in IN 135 138 the the DT 139 151 inflammatory inflammatory JJ 152 159 process process NN 159 160 . . . 161 170 Formation formation NN 171 173 of of IN 174 177 the the DT 178 184 NFAT-1 nfat-1 NN 185 200 transcriptional transcriptional JJ 201 208 complex complex NN 209 211 on on IN 212 215 the the DT 216 220 IL-2 il-2 NN 221 229 promoter promoter NN 230 232 is be VBZ 233 242 essential essential JJ 243 246 for for IN 247 251 IL-2 il-2 NN 252 265 transcription transcription NN 265 266 . . . 267 272 Using use VBG 273 274 a a DT 275 279 cell cell NN 280 284 line line NN 285 289 that that WDT 290 292 is be VBZ 293 299 stably stably RB 300 311 transfected transfecte VBN 312 316 with with IN 317 318 a a DT 319 325 trimer trimer NN 326 328 of of IN 329 332 the the DT 333 339 NFAT-1 nfat-1 NN 340 350 regulatory regulatory JJ 351 358 element element NN 359 365 linked link VBN 366 368 to to TO 369 370 a a DT 371 376 lac-Z lac-z NN 377 385 reporter reporter NN 386 390 gene gene NN 390 391 , , , 392 394 we we PRP 395 403 screened screen VBD 404 407 for for IN 408 418 inhibitors inhibitor NNS 419 421 of of IN 422 437 NFAT-1-mediated nfat-1-mediated JJ 438 456 beta-galactosidase beta-galactosidase NN 457 465 activity activity NN 465 466 . . . 467 470 WIN win NN 471 476 61058 61058 CD 477 480 and and CC 481 484 WIN WIN NNP 485 490 53071 53071 CD 491 495 were be VBD 496 506 identified identify VBN 507 509 as as IN 510 516 microM microm NN 517 527 inhibitors inhibitor NNS 527 528 . . . 529 534 These these DT 535 544 compounds compound NNS 545 549 also also RB 550 559 inhibited inhibit VBD 560 578 beta-galactosidase beta-galactosidase NN 579 583 mRNA mrna NN 584 590 levels level NNS 590 591 . . . 592 599 Similar similar JJ 600 610 inhibition inhibition NN 611 613 of of IN 614 629 NFAT-1-mediated nfat-1-mediated JJ 630 634 gene gene NN 635 645 expression expression NN 646 649 was be VBD 650 658 observed observe VBN 659 661 in in IN 662 663 a a DT 664 670 second second JJ 671 675 cell cell NN 676 680 line line NN 680 681 , , , 682 687 which which WDT 688 690 is be VBZ 691 697 stably stably RB 698 709 transfected transfecte VBN 710 714 with with IN 715 721 NFAT-1 nfat-1 NN 722 732 regulatory regulatory JJ 733 741 elements element NNS 742 748 linked link VBN 749 751 to to TO 752 755 the the DT 756 764 reporter reporter NN 765 769 gene gene NN 770 773 for for IN 774 778 sCD8 scd8 NN 778 779 . . . 780 782 At at IN 783 785 10 10 CD 786 792 microM microM NNP 792 793 , , , 794 798 both both DT 799 808 compounds compound NNS 809 818 inhibited inhibit VBD 819 823 IL-2 il-2 NN 824 828 mRNA mrna NN 829 832 and and CC 833 840 protein protein NN 841 847 levels level NNS 848 850 in in IN 851 854 the the DT 855 868 NFAT-1-linked nfat-1-linked JJ 869 874 lac-Z lac-z NN 875 888 transfectants transfectant NNS 888 889 , , , 890 893 and and CC 894 896 in in IN 897 902 human human JJ 903 914 lymphocytes lymphocyte NNS 914 915 . . . 916 920 Both both DT 921 930 compounds compound NNS 931 940 inhibited inhibit VBD 941 944 the the DT 945 950 mixed mixed JJ 951 961 lymphocyte lymphocyte NN 962 970 reaction reaction NN 970 971 , , , 972 975 and and CC 976 980 this this DT 981 991 inhibition inhibition NN 992 995 was be VBD 996 1004 reversed reverse VBN 1005 1007 by by IN 1008 1017 exogenous exogenous JJ 1018 1022 IL-2 il-2 NN 1022 1023 . . . 1024 1027 WIN win NN 1028 1033 53071 53071 CD 1034 1043 inhibited inhibit VBD 1044 1048 IL-2 il-2 NN 1049 1059 production production NN 1060 1067 induced induce VBN 1068 1070 in in IN 1071 1074 the the DT 1075 1092 calcium-dependent calcium-dependent JJ 1093 1096 PMA pma NN 1097 1100 and and CC 1101 1110 ionomycin ionomycin NN 1111 1118 pathway pathway NN 1118 1119 . . . 1120 1130 Conversely conversely RB 1130 1131 , , , 1132 1151 calcium-independent calcium-independent JJ 1152 1161 anti-CD28 anti-cd28 JJ 1162 1164 Ab Ab NNP 1165 1168 and and CC 1169 1180 PMA-induced pma-induced JJ 1181 1185 IL-2 il-2 NN 1186 1196 production production NN 1197 1200 was be VBD 1201 1210 resistant resistant JJ 1210 1211 . . . 1212 1216 Both both DT 1217 1226 compounds compound NNS 1227 1234 altered alter VBD 1235 1238 the the DT 1239 1245 NFAT-1 nfat-1 NN 1246 1261 transcriptional transcriptional JJ 1262 1269 complex complex NN 1269 1270 , , , 1271 1278 causing cause VBG 1279 1282 its its PRP$ 1283 1291 retarded retard VBN 1292 1300 mobility mobility NN 1301 1303 on on IN 1304 1308 gels gel NNS 1308 1309 . . . 1310 1312 By by IN 1313 1318 these these DT 1319 1329 functional functional JJ 1330 1338 criteria criterion NNS 1338 1339 , , , 1340 1342 we we PRP 1343 1350 believe believe VBP 1351 1353 we we PRP 1354 1358 have have VBP 1359 1369 identified identify VBN 1370 1373 two two CD 1374 1386 structurally structurally RB 1387 1395 distinct distinct JJ 1395 1396 , , , 1397 1402 novel novel JJ 1403 1413 inhibitors inhibitor NNS 1414 1416 of of IN 1417 1432 NFAT-1-mediated nfat-1-mediated JJ 1433 1446 transcription transcription NN 1446 1447 . . .