Expression analysis of DAPK and IRF-1 in primitive leukemic and normal cells.

(A) RT-PCR was performed on the equivalent of 1000 purified cells from normal bone marrow (BM) or AML-derived CD34⁺/CD38⁻ cells. Lanes 1-3 are independent normal marrow CD34⁺/CD38⁻ cell specimens. The remaining 4 lanes are derived from independent AML blood specimens sorted to isolate CD34⁺/CD38⁻ populations. PCRs for IRF-1 and DAPK are shown in the top and middle panels, respectively. The bottom panel shows control reactions using primers for β2-microglobulin. (B) Immunoblot analysis was performed to analyze IRF-1 and DAPK with the use of the equivalent of 400 000 cells per lane (each lane is an independent specimen). Samples were derived from flow-cytometrically sorted normal CD34⁺ cells (first 4 lanes after the KG-1 cell line control) or sorted AML CD34⁺cells (adjacent 9 lanes). From left to right, AML specimens were from FAB types M4, M1, M1, M2/6, M2, M4, M4, M5, and M4.