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. 2016 Aug 25;7(40):65732–65743. doi: 10.18632/oncotarget.11610

Figure 2. Activation of p53 signaling is responsible for cell cycle arrest and apoptosis in response to GCM.

Figure 2

A. GCM activated p53 signaling pathway in HepG2 cells. The cells were exposure to various concentrations of GCM for 24 h and the changes of p-p53, p21 and puma were analyzed by western blotting. B.&C. Effects of p53 inhibition by RNAi on GCM-induced cell death and cell cycle arrest in HepG2 cells. The cells were transfected with 7.5 nmol/L of p53 siRNA or non-targeting siRNA for 24 h and then treated with GCM for 24 h. Cell death was measured by Annexin V/FITC Staining (B) and cell cycle distribution was assessed by flow cytometry following staining with propidium iodide (C). D. GCM induced p53-dependent up-regulation of p21 and puma in HCT-116 cells. HCT-116 p53+/+ or p53−/− cells were treated with various concentrations of GCM for 24 h and the changes of p-p53, p21 and puma were analyzed by western blotting. E. Overall inhibitory effects of GCM on HCT-116 p53+/+ or p53−/− cells measured by crystal violet staining. F. Effects of GCM on cell cycle distribution in HCT-116 p53+/+ or p53−/− cells measured by flow cytometry following staining with propidium iodide. G. Apoptosis induction by GCM in HCT-116 p53+/+ or p53−/− cells analyzed by Annexin V/FITC Staining.