Figure 2

Par-4 cleavage is a common step in the process of caspase-dependant apoptosis. Panel A , Jurkat cells (0.2 μM Dox, 4 μM Eto and 25 μM Cur), MCF-7 cells (3 μM Dox, 100 μM Eto and 40 μM Cur) and LNCaP cells (5 μM Dox, 50 μM Eto and 50 μM Cur) were treated for 24 h and cleavage of Par-4 was analyzed by western blot analysis using Par-4 antibody. Apoptosis induction was confirmed by PARP cleavage and actin was used as protein loading control. Panel B , Jurkat, MCF-7 and LNCaP cells were treated with Dox, Eto and Cur for 24 h and cell viability was measured by using WST viability assay kit. Data shown are means ± SD (n = 3). *, p < 0.05 compared with untreated control. Panel C , MCF-7 cells were treated with indicated concentration of Dox and intracellular SPH generation was measured. Data shown are means ± SD (n = 3).*, p < 0.05 compared with untreated control. Panel D , MCF-7 cells were treated with indicated concentration of Dox, cells were lysed and Western blot analysis of Par-4 and PARP were carried out. Actin was used as a loading control. Panel E , MCF-7 cells were transiently transfected with control siRNA (Cont siRNA) and Par-4 siRNA. After 24 h, cells were treated with either vehicle or 3 μM Dox for further 24 h. The expression of Par-4 and PARP were detected by using Western blot analysis. Actin was used as the protein loading control. Panel F , MCF-7 cells were transiently transfected with Cont siRNA and Par-4 siRNA. After 24 h cells were treated with either vehicle or 3 μM Dox for further 24 h. The cell viability assay was performed. Data shown are means ± SD (n = 3). *, p < 0.05, and **, p < 0.05 compared with untreated control and control siRNA Dox treated cells, respectively.