Immunoblot analysis of Pim target proteins in AML cell line treated with SGI-1776. MV-4-11 cells were either untreated, treated with 0.1% vehicle DMSO alone, 0.1, 0.3, 1, 3, or 10μM SGI-1776 and cells were harvested after 24 hours and lysed. (A) The phospho-protein levels of histone H3 (Ser10), c-Myc (Ser62), Bad(Ser112), 4E-BP1 (Thr37/46), were analyzed using immunoblot and normalized to each respective total protein levels. (B) Phospho-protein levels relative to total protein levels. Data are the average of 3 independent experiments ± SEM. (C) Protein levels of FLT3 and p27 were analyzed using immunoblot and normalized to GAPDH as a loading control. (D) p27 protein levels normalized to GAPDH protein. Data are the average of 3 independent experiments ± SEM. (E) Protein levels of proapoptotic Bak, Bax, and antiapoptotic Mcl-1, Bcl-2, and Bcl-x_L were analyzed using immunoblot and normalized to GAPDH as a loading control. (F) Pro- and antiapoptotic protein levels normalized to GAPDH protein with Mcl-1 shown in gray. Data are the average of 3 independent experiments ± SEM.