Figure 4.

Type B response pattern in birinapant-sensitive WK1luc cells. (A–D) Flow cytometry analysis demonstrated that birinapant and birinapant plus TMZ induced significant zVAD-sensitive (‘secondary necrotic') cell death. (E, F) A similar finding was observed by Hoechst staining; data are shown as a boxplot with overlaid data points from at least three independent experiments with technical replicates (two-way ANOVA, ***P<0.001 versus control cells, ⁺⁺⁺P<0.001 versus treated cells. Scale bar, 50 μm. (G) Western blot analysis indicated that birinapant treatment readily activated caspase-8, and lead to an increase in FADD and reduction in RIP1 protein levels. Actin was used as a loading control.