Cytotoxic effect of PKC412 on primary AML cells leads to up-regulation of Bim and Puma. Mononuclear cells were prepared from cryopreserved bone marrow or peripheral blood samples of 4 AML patients. (A) Apoptosis assay was performed with PKC412 at 2 high concentrations (100 and 300 nM) on AML cells cultured for 72 hours, then stained with PI, and analyzed by flow cytometry. Data are mean (± SD) from duplicates. Symbols indicate patients as: ■, AML no. 1; □, AML no. 3; ♦, AML no. 2; ◇, AML no. 4. (B) The relative expression levels of Bim (□) and Puma () mRNA from the same AML patients were measured by real-time PCR after 24 hours of treatment with 30 and 100 nM PKC412. Results are from triplicate reactions.