Influence of sCD40L at different concentrations on the expression of Bcl-2 family members.
L3055 (wild-type) cells at 2 × 105 per mL were cultured for 3 days with concentrations of sCD40L indicated. (Upper left panel) Lysates were prepared and equal protein loadings (50 μg) subjected to Western blotting for Bcl-2 protein on 12.5% gels as described in “Materials and methods”; molecular weight markers are indicated to the right. (Right panels) Cells were harvested and stained for the expression of Bcl-2 (upper), Bcl-xL (middle), Bax (lower) and analyzed by FACS as described in “Materials and methods.” Results are given as means (with SE) of 3 separate experiments with percentage of cells positive shown as histograms and the mean fluorescence intensities (MFI) of positively gated cells as symbols. (Lower left panel) The levels of Bcl-2, Bcl-xL, and Bax expression in L3055 cells transfected with bcl-2 as determined by FACS analysis.
