Figure 2

Expression of apoptosis regulatory genes in MLL-AF9 AMLs. (a) Western blot and (b) qPCR analysis of expression of p53, p19Arf and bcl-2 gene family members in bone marrow cells of sick primary MLL-AF9 AML mice (Supplementary Methods). Protein and RNA were prepared from bone marrow taken at autopsy. Panels in a indicate six separate gels, each of which included WT AML #1300, with dotted lines separating different genotypes (the solid line in panel 3 indicates electronic removal of a lane). SV40 transformed mouse embryonic fibroblasts (MEF) served as positive controls for p53 and p19Arf expression; Hsp70 served as a loading control. Molecular weight (kD) markers are indicated. The following AMLs were analyzed by qPCR: WT #1223, 1224, 1155, 1156, 1231, 1232, 1601, 1410, 1411, 1414; bim^−/− #1249, 1250, 1213, 1157, 1158; puma^−/− #1230, 1214, 1215, 1216, 1217, 1218, 1225, 1226, 1228, 1229; noxa^−/− #1259, 1313, 1308, 1310; bmf^−/− #1440, 1441, 1442, 1416, 1419, 1418; puma^−/− bim^−/− # 1590, 1593; noxa^−/− bmf ^−/− #1596; 1597; 1598; puma^−/− noxa^−/− #1443; 1444, 1445. qPCR analysis was relative to that of hydroxymethylbilane synthase (HMBS). Error bars indicate S.D. Data for additional Bcl-2 family members is provided in Supplementary Figure S4