Figure 1.

Bufalin-activated AKT in MM cells and the combination of bufalin with MK2206 increased the rate of apoptosis induction. (a) H929 and U266 cells were incubated with 12 nM of bufalin for 3, 6, 12 h and subsequently subjected to western blot analysis using anti-p-AKT, anti-AKT and anti-β-actin antibodies. β-actin was used as a loading control. (b) H929 and U266 cells were treated with 12 nM of bufalin and/or 6 μM of MK2206 for 48 h and the apoptotic rates were analyzed by flow cytometry. The apoptotic effect of the combination group was statistically different compared with treatment of bufalin and/or MK2206 alone. Each bar represented the mean±S.E. of triplicate experiments. The values under the bands represented the mean quantitation ratios compared with the control groups. (c) H929 and U266 cells were treated with 12 nM of bufalin in the absence and/or presence of 6 μM of MK2206 for 12, 24, 36, 48 h and the protein lysates were subjected to immunoblot analysis using antibodies specific against PARP, caspase-3, caspase-9 and/or β-actin. β-actin was used as a loading control. Experiments were performed in triplicate (*P<0.05; **P<0.01)