Fig. 3. Role of p53 in cell cycle arrest after leflunomide treatment.
a MCF7 and b HCT116 cells were exposed to LFM (50 µM) for 72 h after downregulation of RPL5 and RPL11 alone or in combination using specific siRNA. Cell death was assessed by flow cytometry using annexin V/Hoechst staining. Non-targeting siRNA (siNC) was used as a control. c MCF7 cells were transfected with specific p53 siRNA or with non-targeting siRNA (siNC), respectively. Cell death was evaluated by annexin V/Hoechst positivity using FACS. Immunoblot detection of p53 and p21 shows the effect of transfection. β-Actin was used as a loading control. d HCT116 wt p53 and HCT116 p53KO cells were exposed to LFM (50 µM) for 72 h and cell death was determined by flow cytometry using annexin V/Hoechst staining. Immunoblot detection of p53 and p21 show the effect of p53KO. β-Actin was used as a loading control. In a–d, data are shown as mean ± SEM, n = 3-5. *P < 0.05, two-way ANOVA.