LBH589 alters the level of intact mTORC2 and c-Myc expression. (A-B) Effect of siRNA knockdown of raptor or rictor. (A) Rictor siRNA blocks phosphorylation of Akt in the presence of rapamycin. Ly7 cells were transiently transfected with raptor or rictor siRNA and then treated with rapamycin (R) or left untreated for 24 hours and subjected to Western blot analysis using p-Akt antibody. (B) After raptor or rictor knockdown, DHL-6 cells were treated with rapamycin (R) or left untreated (0) for 24 hours. Survival was analyzed by staining with annexin V and PI. (C-D) Effect of LBH and rapamycin on cyclin D1 and c-Myc expression. (C) Ly7 cells were treated with the indicated concentrations of LBH589 or rapamycin for 24 hours and then subjected to Western blot analysis using c-Myc and cyclin D1 antibodies. (D) Ly cells were cotreated with 50 nM LBH and rapamycin for 24 hours, and c-Myc and cyclin D1 expression was analyzed. (E-F) DHL-6 cells were transiently transfected with control siRNA or c-Myc siRNA, and proliferation was done as described earlier. Western blot analysis showed successful knockdown of c-Myc (50 nM or 200 nM). (G) HDAC3-transfected DHL-6 cells were analyzed for c-Myc and cyclin D1 expression. Error bars indicate mean ± SD from 3 determinations.