Helenalin induced caspase cleavage and autophagy is dependent on Atg12 and LC3-B. A2780 cells were transfected with either non-targeting siRNA (siRNA Neg) or Atg12 or LC3-B-specific siRNA for 24 h and then treated with either DMSO or helenalin for 24 h. Cells were harvested and (A and B) cell lysates were subjected to immunoblot analysis for Atg12, LC3-B, cleaved PARP, caspase 3 or 9 or (C and D) analyzed for cell cycle phase distribution by FACS. All experiments were performed in biological triplicates. Actin was used as the loading control. One way ANOVA was performed between control and treatment groups, and significant differences were observed between the groups as indicated in Figure 6C and D. Categories significant after multiple comparison are marked as “Yes”.(p-value <0.05; Tukey’s multiple comparison test).