Induction of activated Akt markedly protect cells from apoptosis induced by coadministration of 2-ME and the HDACIs, NaB and SAHA. U937 cells were stably transfected with constitutively active forms of Akt (clone designated CA-3) or an empty vector (pUSE) as described in “Materials and methods.” Cells were then treated with 0.5 μM 2-ME in the presence or absence of NaB (1 mM) or SAHA (1.5 μM) for 24 hours. (A) After treatment, apoptosis was analyzed using annexin V-FITC/PI assay as described in “Materials and methods.” ^*Values for Akt-CA3 cells treated with 2-ME in the presence of NaB or SAHA were significantly decreased compared with those for pUSE cells (Student t test; P < .01). (B) After treatment, total cellular or cytosolic extracts were prepared and subjected to Western blot analysis using antibodies against PARP, caspase-8, caspase-9, cleaved caspase-3, AIF, and cytochrome c, respectively. (C) After treatment, total cellular extracts were prepared and subjected to Western blot analysis using antibodies against phosphor-Akt, Akt, phospho-JNK, and JNK. For Western blot analysis, each lane was loaded with 30 μg protein. Blots were subsequently stripped and reprobed with antibody against β-actin to ensure equivalent loading and transfer. Two additional studies yielded equivalent results.