0 13 Transcription Transcription NNP 14 21 factors factor NNS 22 25 Sp1 sp1 NN 26 29 and and CC 30 34 AP-2 ap-2 NN 35 42 mediate mediate VBP 43 52 induction induction NN 53 55 of of IN 56 60 acid acid NN 61 77 sphingomyelinase sphingomyelinase NN 78 84 during during IN 85 94 monocytic monocytic JJ 95 110 differentiation differentiation NN 110 111 . . . 113 118 Cells cell NNS 119 123 from from IN 124 127 the the DT 128 133 human human JJ 134 143 monocytic monocytic JJ 144 152 leukemia leukemia NN 153 157 cell cell NN 158 162 line line NN 163 168 THP-1 thp-1 NN 169 182 differentiate differentiate VBP 183 190 towards towards IN 191 192 a a DT 193 208 macrophage-like macrophage-like JJ 209 218 phenotype phenotype NN 219 223 when when WRB 224 234 stimulated stimulate VBN 235 239 with with IN 240 247 phorbol phorbol NN 248 260 12-myristate 12-myristate NN 261 265 -13- -13- NN 266 273 acetate acetate NN 274 275 ( ( ( 275 278 PMA PMA NNP 278 279 ) ) ) 279 280 , , , 281 303 1,25-dihydroxy-vitamin 1,25-dihydroxy-vitamin NN 304 306 D3 d3 NN 306 307 , , , 308 311 and and CC 312 319 various various JJ 320 325 other other JJ 326 332 agents agent NNS 332 333 . . . 334 336 We we PRP 337 348 demonstrate demonstrate VBP 349 353 here here RB 354 358 that that IN 359 362 the the DT 363 373 expression expression NN 374 376 of of IN 377 380 the the DT 381 390 lysosomal lysosomal JJ 391 397 enzyme enzyme NN 398 402 acid acid NN 403 419 sphingomyelinase sphingomyelinase NNP 420 421 ( ( ( 421 424 ASM ASM NNP 424 425 ; ; : 426 438 E.C.3.1.4.12 E.C.3.1.4.12 NNP 438 439 ) ) ) 440 442 is be VBZ 443 450 induced induce VBN 451 457 during during IN 458 462 this this DT 463 470 process process NN 471 474 and and CC 475 477 is be VBZ 478 486 strongly strongly RB 487 495 elevated elevated JJ 496 498 in in IN 499 513 differentiated differentiate VBN 514 519 THP-1 thp-1 NN 520 525 cells cell NNS 525 526 , , , 527 529 as as RB 530 534 well well RB 535 537 as as IN 538 540 in in IN 541 555 differentiated differentiate VBN 556 561 human human JJ 562 573 mononuclear mononuclear JJ 574 584 phagocytes phagocyte NNS 584 585 . . . 586 591 Using use VBG 592 600 Northern northern JJ 601 609 blotting blotting NN 609 610 , , , 611 616 RNase RNase NNP 617 627 protection protection NN 628 633 assay assay NN 633 634 , , , 635 638 and and CC 639 646 nuclear nuclear JJ 647 653 run-on run-on JJ 654 662 analyses analysis NNS 662 663 , , , 664 666 we we PRP 667 671 show show VBP 672 676 that that IN 677 680 the the DT 681 694 up-regulation up-regulation NN 695 697 of of IN 698 701 ASM ASM NNP 702 712 expression expression NN 713 715 is be VBZ 716 725 regulated regulate VBN 726 732 mainly mainly RB 733 735 at at IN 736 739 the the DT 740 745 level level NN 746 748 of of IN 749 762 transcription transcription NN 763 766 and and CC 767 771 that that IN 772 775 new new JJ 776 783 protein protein NN 784 793 synthesis synthesis NN 794 796 is be VBZ 797 805 required require VBN 806 809 for for IN 810 818 enhanced enhance VBN 819 822 ASM ASM NNP 823 831 activity activity NN 831 832 . . . 833 837 This this DT 838 847 cell-type cell-type JJ 848 856 specific specific JJ 857 866 induction induction NN 867 869 by by IN 870 873 PMA PMA NNP 874 883 treatment treatment NN 884 887 was be VBD 888 895 further further RBR 896 908 investigated investigate VBN 909 913 with with IN 914 921 respect respect NN 922 924 to to TO 925 940 transcriptional transcriptional JJ 941 948 control control NN 948 949 . . . 950 951 A a DT 952 958 series series NN 959 961 of of IN 962 963 5 5 CD 963 964 ’ ' SYM 965 973 deletion deletion NN 974 985 derivatives derivative NNS 986 988 of of IN 989 992 the the DT 993 1001 upstream upstream JJ 1002 1012 regulatory regulatory JJ 1013 1019 region region NN 1020 1024 were be VBD 1025 1029 used use VBN 1030 1032 in in IN 1033 1042 transient transient JJ 1043 1055 transfection transfection NN 1056 1062 assays assay NNS 1063 1065 to to TO 1066 1074 identify identify VB 1075 1083 promoter promoter NN 1084 1092 elements element NNS 1093 1101 required require VBN 1102 1105 for for IN 1106 1111 basal basal JJ 1112 1115 and and CC 1116 1125 inducible inducible JJ 1126 1130 gene gene NN 1131 1141 expression expression NN 1141 1142 . . . 1143 1144 A a DT 1145 1148 PMA PMA NNP 1149 1159 responsive responsive JJ 1160 1167 element element NN 1168 1171 was be VBD 1172 1181 localized localized JJ 1182 1184 to to TO 1185 1186 a a DT 1187 1193 region region NN 1194 1201 between between IN 1202 1206 -319 -319 CD 1207 1210 and and CC 1211 1215 -219 -219 CD 1216 1218 bp bp NN 1219 1227 upstream upstream RB 1228 1230 of of IN 1231 1234 the the DT 1235 1245 initiation initiation NN 1246 1251 codon codon NN 1252 1255 and and CC 1256 1272 co-transfections co-transfection NNS 1273 1277 with with IN 1278 1291 transcription transcription NN 1292 1298 factor factor NN 1299 1309 expression expression NN 1310 1318 plasmids plasmid NNS 1319 1322 for for IN 1323 1327 AP-2 ap-2 NN 1328 1331 and and CC 1332 1335 Sp1 Sp1 NNP 1336 1344 resulted result VBD 1345 1347 in in IN 1348 1357 augmented augmented JJ 1358 1361 ASM ASM NNP 1362 1370 promoter promoter NN 1371 1379 activity activity NN 1379 1380 , , , 1381 1386 which which WDT 1387 1390 was be VBD 1391 1400 abolished abolish VBN 1401 1405 when when WRB 1406 1409 the the DT 1410 1417 binding bind VBG 1418 1423 sites site NNS 1424 1427 for for IN 1428 1433 these these DT 1434 1437 two two CD 1438 1445 factors factor NNS 1446 1450 were be VBD 1451 1458 deleted delete VBN 1458 1459 . . . 1460 1465 Using use VBG 1466 1481 electrophoretic electrophoretic JJ 1482 1490 mobility mobility NN 1491 1496 shift shift NN 1497 1503 assays assay NNS 1504 1507 and and CC 1508 1518 supershift supershift NN 1519 1525 assays assay NNS 1526 1528 we we PRP 1529 1540 demonstrate demonstrate VBP 1541 1545 that that IN 1546 1550 this this DT 1551 1557 region region NN 1558 1560 is be VBZ 1561 1573 specifically specifically RB 1574 1579 bound bind VBN 1580 1582 by by IN 1583 1586 Sp1 Sp1 NNP 1587 1590 and and CC 1591 1595 AP-2 ap-2 NN 1595 1596 . . . 1597 1602 These these DT 1603 1610 factors factor NNS 1611 1614 are be VBP 1615 1622 present present JJ 1623 1625 in in IN 1626 1633 nuclear nuclear JJ 1634 1642 extracts extract NNS 1643 1651 prepared prepare VBN 1652 1656 from from IN 1657 1661 both both CC 1662 1669 induced induce VBN 1670 1673 and and CC 1674 1683 uninduced uninduced JJ 1684 1689 THP-1 thp-1 NN 1690 1695 cells cell NNS 1695 1696 . . . 1697 1704 However however RB 1704 1705 , , , 1706 1709 the the DT 1710 1719 intensity intensity NN 1720 1722 of of IN 1723 1726 the the DT 1727 1734 complex complex NN 1735 1741 formed form VBN 1742 1750 appeared appear VBD 1751 1753 to to TO 1754 1762 increase increase VB 1763 1767 when when WRB 1768 1775 nuclear nuclear JJ 1776 1784 extracts extract NNS 1785 1789 from from IN 1790 1801 PMA-treated pma-treated JJ 1802 1807 cells cell NNS 1808 1812 were be VBD 1813 1817 used use VBN 1817 1818 . . . 1819 1823 From from IN 1824 1829 these these DT 1830 1837 studies study NNS 1837 1838 , , , 1839 1841 we we PRP 1842 1850 conclude conclude VBP 1851 1855 that that IN 1856 1857 a a DT 1858 1867 concerted concerted JJ 1868 1874 action action NN 1875 1877 of of IN 1878 1881 the the DT 1882 1895 transcription transcription NN 1896 1903 factors factor NNS 1904 1908 AP-2 ap-2 NN 1909 1912 and and CC 1913 1916 Sp1 Sp1 NNP 1917 1919 is be VBZ 1920 1929 essential essential JJ 1930 1933 for for IN 1934 1937 the the DT 1938 1940 up up JJ 1941 1952 -regulation -regulation NN 1953 1955 of of IN 1956 1959 ASM ASM NNP 1960 1970 expression expression NN 1971 1977 during during IN 1978 1981 the the DT 1982 1989 process process NN 1990 1992 of of IN 1993 2003 macrophage macrophage NN 2004 2019 differentiation differentiation NN 2019 2020 . . .