by western blot and spectrofuorometric technique.
A: Western blot analysis showing cleavage of procaspase-8 in untreated control cells (lane 1), cells treated with 5 mM (lane 2) benzamide for 20 h. The un-cleaved band of 55 kDa in lane 1 represents the procaspase-8, while in lane 2 there was cleaved product with 26 kDa and 18 kDa (the active caspase-8). B: Graphic representation of caspase-8 activity in K562 cells treated with different concentrations of benzamide for 20 h (inset) and 5 mM benzamide for various time intervals evaluated using fluorescence assay kit, as described in methods. Each histogram represents the mean ± S.D. of caspase-8 activity obtained from four 30 independent experiments and p-values, calculated using ANOVA for each concentration and time point, indicating the level of significance compared with untreated control (or 0 h for time kinetics).
