Figure 6. ABT-263 induces activation of ERK and survivin up-regulation in HCC cells.

A. LH86 cells were treated with different doses of ABT-263 as indicated for 1 h. Cells were harvested and cell lysates were prepared for Western blotting. Phospho-ERK and ERK protein levels were examined with specific antibodies. β-actin was assessed and served as an equal protein loading control. B, LH86 cells were untreated or treated with ABT-263 (1 µM) for 1 h. Cells were harvested and cell lysates were prepared for Western blotting. Survivin expression level was examined with specific antibodies. β-actin was assessed and served as an equal protein loading control. C. LH86 cells were not treated (control) or treated with ABT-263(1 µM), ERK specific inhibitor PD98059 (50 µM), or pre-treated with PD98059 (50 µM) for 1 h followed by ABT-263 (1 µM) for 24 h. Apoptosis was determined by Hoechst staining to show cells with apoptotic nuclei (representative apoptotic cells were labeled with white arrows; PD: PD98059). D. LH86 cells were treated as in Figure 6C, apoptosis was assessed by nuclear staining and cells with apoptotic nuclei were counted as described in ‘materials and methods’ (*p<0.05). E. LH86 cells were untransfected or transiently transfected with synthesized random siRNA (control) or ERK specific siRNA duplexes, and 48 h post-transfection, cells were subjected to Western blotting analysis with anti-ERK polyclonal antibody. β-actin was used as an equal protein loading control. F. LH86 cells were transfected with synthesized random control siRNA or ERK specific siRNA, and 48 h post-transfection, cells were untreated or treated with ABT-263 (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).