Figure 6.
Effect of sst2 transfection on mitochondrial Bcl-2 expression and cytochrome c localization in BxPC-3 cells. (a and b) Mock- and sst2-transfected BxPC-3 cells were grown (105 cells per ml) and starved for 24 h. (a) For immunoblotting, an anti-Bcl-2 antibody was used, and histone expression was utilized as an internal loading control (n = 3). (b) An immunoblot of cytosolic cytochrome c and histone expression in cytosolic extracts was used as an internal loading control. Expression of COXII was used as a cytosolic extract purity control (n = 3). (c) MTT assay. Mock- and sst2-transfected BxPC-3 cells were grown (104 cells per well) and treated for 24 h with TNFα + cycloheximide, TRAIL, or anti-CD95 Ab. Untreated cells were used as controls. Mock-transfected BxPC-3 cells were also treated (mock + HA14-1) or not treated (mock) with 50 μM Bcl-2 inhibitor HA14-1. Data points represent the mean ± SEM (n = 4) and are expressed as the percentage of cell viability observed in untreated mock-transfected cells (control mock). *, P < 0.05; **, P < 0.01 for HA14-1-treated mock-transfected cells and sst2- vs. mock-transfected cells.