Effect of AZD1208 on cell growth and survival in AML cell lines. (A) Pim-1 and Pim-2 expression and STAT3 and STAT5 expression and phosphorylation across a panel of AML cell lines with varying sensitivity to AZD1208, as shown by immunoblot analysis of cell lysates and GI₅₀ values from a 72-hour proliferation assay. (B) AZD1208 effect on G₁ cell cycle arrest and subG₁ population in MOLM-16 cells, assessed by propidium iodide staining and flow cytometry for DNA content 72 hours after treatment. Data shown are representative of 3 independent experiments. (C) Effect of AZD1208 on apoptosis in MOLM-16 cells determined by staining with Annexin V (early apoptosis), Annexin V and 7-amino-actinomycin D (7-AAD) (late apoptosis), and 7-AAD (dead/necrotic cells). Data shown are representative of 3 independent experiments. (D) AZD1208 effect on p27, pBAD, and cleaved caspase 3, as shown by immunoblot anaylsis 3 and 24 hours after treatment. FL2-H, peak emission values in the FL2 channel; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; ND, not determined.