Figure 5. Bcl-xL down-regulation enhances YM-155-induced apoptotic toxicity in HCC cells.

A. Huh7 cells were transiently transfected with synthesized random siRNA (control) or Bcl-xL specific siRNA duplexes, and 48 h post-transfection, cells were subjected to Western blotting analysis with anti-Bcl-xL polyclonal antibody. β-actin was used as an equal protein loading control. B. Huh7 cells were transfected with synthesized random control siRNA or Bcl-xL specific siRNA, and 48 h post-transfection, cells were untreated or treated with YM-155 (1 µM) for 24 h and then subjected to Hoechst staining to show apoptotic cells with condensed nuclei (representative apoptotic cells were marked with white arrows). C. Huh7 cells were treated as in with YM-155 as indicated and apoptosis was measured as in Figure 2A. Statistical analysis was performed for apoptosis ratio by counting the number of cells with apoptotic nuclei (*p<0.05). D. Huh7 cells treated were harvested and cell lyates were prepared and subjected to Western blotting. Apoptosis was determined through caspase 3 activation. β-actin was used as an equal protein loading control.