Treatment with AG1295 or PKC412 of human leukemic cell lines expressing mutated FLT3 leads to up-regulation of Bim and Puma and apoptosis induction. Leukemic cell lines expressing no FLT3 (THP1), wild-type FLT3 (NB4, RS4), or mutated FLT3 (MV4;11, MonoMac-6) were seeded at a density of 10⁵-10⁶ cells/mL in the absence or presence of AG1295 or PKC412. (A) Seventy-two hours later, cells were analyzed for apoptosis by flow cytometry after staining with annexin V–FITC/PI. Data shown are mean (± SD) from 3 experiments. *P < .05 (SD over control without inhibitor); **P < .01 (SD); ***P < .001 (SD). (B) Expression of Bim and Puma protein was analyzed by Western blot analysis after 48 hours of treatment. (C) Real-time PCR using Bim and Puma primers from cells after 24 hours of treatment with 30 μM AG1295 (□) or 50 nM PKC412 (). The results are mean (± SD) from 2 experiments performed in triplicates and presented as relative expression compared with the control housekeeping gene GusB.