0 16 Thrombin-induced thrombin-induced JJ 17 20 p65 p65 NN 21 30 homodimer homodimer NN 31 38 binding bind VBG 39 41 to to TO 42 52 downstream downstream JJ 53 61 NF-kappa NF-kappa NNP 62 63 B B NNP 64 68 site site NN 69 71 of of IN 72 75 the the DT 76 84 promoter promoter NN 85 93 mediates mediate VBZ 94 105 endothelial endothelial JJ 106 112 ICAM-1 icam-1 NN 113 123 expression expression NN 124 127 and and CC 128 138 neutrophil neutrophil NN 139 147 adhesion adhesion NN 147 148 . . . 150 152 We we PRP 153 165 investigated investigate VBD 166 169 the the DT 170 180 mechanisms mechanism NNS 181 183 by by IN 184 189 which which WDT 190 205 proinflammatory proinflammatory JJ 206 214 mediator mediator NN 214 215 , , , 216 224 thrombin thrombin NN 224 225 , , , 226 234 released release VBN 235 241 during during IN 242 255 intravascular intravascular JJ 256 267 coagulation coagulation NN 268 271 and and CC 272 278 tissue tissue NN 279 285 injury injury NN 285 286 , , , 287 294 induces induce VBZ 295 301 ICAM-1 ICAM-1 NNP 302 303 ( ( ( 303 307 CD54 cd54 NN 307 308 ) ) ) 309 319 expression expression NN 320 322 in in IN 323 334 endothelial endothelial JJ 335 340 cells cell NNS 340 341 . . . 342 353 Stimulation stimulation NN 354 356 of of IN 357 362 HUVEC HUVEC NNP 363 367 with with IN 368 376 thrombin thrombin NN 377 385 resulted result VBD 386 388 in in IN 389 394 dose- dose- NN 395 398 and and CC 399 413 time-dependent time-dependent JJ 414 423 increases increase NNS 424 426 in in IN 427 433 ICAM-1 icam-1 NN 434 438 mRNA mRNA NNP 439 442 and and CC 443 447 cell cell NN 448 455 surface surface NN 456 466 expression expression NN 467 470 and and CC 471 473 in in IN 474 490 ICAM-1-dependent icam-1-dependent JJ 491 502 endothelial endothelial JJ 503 513 adhesivity adhesivity NN 514 520 toward toward IN 521 538 polymorphonuclear polymorphonuclear JJ 539 549 leukocytes leukocyte NNS 549 550 . . . 551 560 Transient transient JJ 561 573 transfection transfection NN 574 576 of of IN 577 588 endothelial endothelial JJ 589 594 cells cell NNS 595 599 with with IN 600 606 ICAM-1 icam-1 NN 607 615 promoter promoter NN 616 626 luciferase luciferase NN 627 635 reporter reporter NN 636 640 gene gene NN 641 642 ( ( ( 642 651 ICAM-1LUC ICAM-1LUC NNP 651 652 ) ) ) 653 663 constructs construct NNS 664 673 indicated indicate VBD 674 678 that that IN 679 687 deletion deletion NN 688 690 of of IN 691 699 upstream upstream JJ 700 708 NF-kappa NF-kappa NNP 709 710 B B NNP 711 715 site site NN 716 717 ( ( ( 717 721 -533 -533 CD 722 727 bases base NNS 728 732 from from IN 733 744 translation translation NN 745 750 start start NN 751 755 site site NN 755 756 ) ) ) 757 760 had have VBD 761 763 no no DT 764 770 effect effect NN 771 773 on on IN 774 782 thrombin thrombin NN 783 797 responsiveness responsiveness NN 797 798 , , , 799 806 whereas whereas IN 807 824 mutation/deletion mutation/deletion NN 825 827 of of IN 828 838 downstream downstream JJ 839 847 NF-kappa NF-kappa NNP 848 849 B B NNP 850 854 site site NN 855 856 ( ( ( 856 860 -223 -223 CD 861 866 bases base NNS 867 871 from from IN 872 875 the the DT 876 887 translation translation NN 888 893 start start NN 894 898 site site NN 898 899 ) ) ) 900 909 prevented prevent VBD 910 913 the the DT 914 924 activation activation NN 925 927 of of IN 928 934 ICAM-1 icam-1 NN 935 943 promoter promoter NN 943 944 , , , 945 955 indicating indicate VBG 956 960 that that IN 961 964 the the DT 965 975 downstream downstream JJ 976 984 NF-kappa NF-kappa NNP 985 986 B B NNP 987 991 site site NN 992 994 is be VBZ 995 1003 critical critical JJ 1004 1007 for for IN 1008 1016 thrombin thrombin NN 1017 1029 inducibility inducibility NN 1029 1030 . . . 1031 1039 NF-kappa NF-kappa NNP 1040 1050 B-directed b-directed JJ 1051 1061 luciferase luciferase NN 1062 1070 activity activity NN 1071 1080 increased increase VBD 1081 1094 approximately approximately RB 1095 1101 3-fold 3-fold RB 1102 1106 when when WRB 1107 1112 cells cell NNS 1113 1124 transfected transfecte VBN 1125 1129 with with IN 1130 1133 the the DT 1134 1141 plasmid plasmid NN 1142 1151 pNF-kappa pnf-kappa NN 1152 1156 BLUC BLUC NNP 1157 1167 containing contain VBG 1168 1172 five five CD 1173 1179 copies copy NNS 1180 1182 of of IN 1183 1192 consensus consensus NN 1193 1201 NF-kappa NF-kappa NNP 1202 1203 B B NNP 1204 1208 site site NN 1209 1215 linked link VBN 1216 1218 to to TO 1219 1220 a a DT 1221 1228 minimal minimal JJ 1229 1239 adenovirus adenovirus NN 1240 1243 E1B e1b NN 1244 1263 promoter-luciferase promoter-luciferase NN 1264 1268 gene gene NN 1269 1273 were be VBD 1274 1281 exposed expose VBN 1282 1284 to to TO 1285 1293 thrombin thrombin NN 1293 1294 , , , 1295 1305 indicating indicate VBG 1306 1310 that that IN 1311 1321 activation activation NN 1322 1324 of of IN 1325 1333 NF-kappa NF-kappa NNP 1334 1335 B B NNP 1336 1339 was be VBD 1340 1349 essential essential JJ 1350 1353 for for IN 1354 1362 thrombin thrombin NN 1363 1371 response response NN 1371 1372 . . . 1373 1376 Gel Gel NNP 1377 1387 supershift supershift NN 1388 1394 assays assay NNS 1395 1407 demonstrated demonstrate VBD 1408 1412 that that IN 1413 1421 thrombin thrombin NN 1422 1429 induced induce VBD 1430 1437 binding binding NN 1438 1440 of of IN 1441 1449 NF-kappa NF-kappa NNP 1450 1454 Bp65 Bp65 NNP 1455 1456 ( ( ( 1456 1459 Rel Rel NNP 1460 1461 A A NNP 1461 1462 ) ) ) 1463 1465 to to TO 1466 1476 downstream downstream JJ 1477 1485 NF-kappa NF-kappa NNP 1486 1487 B B NNP 1488 1492 site site NN 1493 1495 of of IN 1496 1499 the the DT 1500 1506 ICAM-1 icam-1 NN 1507 1515 promoter promoter NN 1515 1516 . . . 1517 1525 Thrombin Thrombin NNP 1526 1534 receptor receptor NN 1535 1545 activation activation NN 1546 1553 peptide peptide NN 1553 1554 , , , 1555 1556 a a DT 1557 1570 14-amino-acid 14-amino-acid JJ 1571 1578 peptide peptide NN 1579 1591 representing represent VBG 1592 1595 the the DT 1596 1599 new new JJ 1600 1603 NH2 nh2 NN 1604 1612 terminus terminus NN 1613 1615 of of IN 1616 1631 proteolytically proteolytically RB 1632 1641 activated activate VBN 1642 1652 receptor-1 receptor-1 NN 1652 1653 , , , 1654 1662 mimicked mimic VBD 1663 1671 thrombin thrombin NN 1671 1673 ’s 's POS 1674 1680 action action NN 1681 1683 in in IN 1684 1692 inducing induce VBG 1693 1699 ICAM-1 icam-1 NN 1700 1710 expression expression NN 1710 1711 . . . 1712 1717 These these DT 1718 1722 data datum NNS 1723 1731 indicate indicate VBP 1732 1736 that that IN 1737 1745 thrombin thrombin NN 1746 1755 activates activate VBZ 1756 1767 endothelial endothelial JJ 1768 1774 ICAM-1 icam-1 NN 1775 1785 expression expression NN 1786 1789 and and CC 1790 1807 polymorphonuclear polymorphonuclear JJ 1808 1817 leukocyte leukocyte NN 1818 1826 adhesion adhesion NN 1827 1829 by by IN 1830 1838 NF-kappa NF-kappa NNP 1839 1843 Bp65 Bp65 NNP 1844 1851 binding bind VBG 1852 1854 to to TO 1855 1858 the the DT 1859 1869 downstream downstream JJ 1870 1878 NF-kappa NF-kappa NNP 1879 1880 B B NNP 1881 1885 site site NN 1886 1888 of of IN 1889 1895 ICAM-1 icam-1 NN 1896 1904 promoter promoter NN 1905 1910 after after IN 1911 1926 proteolytically proteolytically RB 1927 1936 activated activate VBN 1937 1947 receptor-1 receptor-1 JJ 1948 1958 activation activation NN 1958 1959 . . .