Fig. 7.

Nutlin-3 + THZ1 potentiated GSDME cleavage. A Western blot was meant to detect GSDME-N protein levels in MCF-7 cell which was treated with 50 M THZ1and 20uM nutlin-3 for 48 h. B Quantity of GSDME-N in A. C The release of LDH was measured after MCF-7 cells treated with 50 nM THZ1 + 20uM nutlin-3 for 48 h. D Western blot was meant to detect GSDME-N protein levels in MCF-7 cell which was treated with 50 nM THZ1 for 48 h after WT p53 plasmids transfection. E Quantity of GSDME-N in D. F The release of LDH was measured after MCF-7 cells treated with 50 nM THZ1 for 48 h after WT p53 plasmids transfection. G MCF-7 cell was treated with 50 nM THZ1 combining increasing concentrations of doxorubicin for 48H. The protein level of GSDME-N was detected by western blot. H Quantity of GSDME-N in G. I MCF-7 cell was co-incubated with ZVAD for 6 h, then treated with 50 nM THZ1 + 20uM nutlin-3 for 48H. The protein level of GSDME-N was detected by western blot. J Quantity of GSDME-N in I. K The release of LDH of MCF-7 cell which firstly incubated with ZVAD for 6 h, then treated with 50 nM THZ1 + 20uM nutlin-3 for 48H. L MCF-7 cell was firstly given ZVAD for 6 h, then treated with 50 M THZ1and 20uM nutlin-3. CCK8 assay was used to measure the 24 h or 48 h cell viability after combination was added. M Colony formation ability measurement of MCF-7 cell after a 6 h ZVAD pretreatment, then added with 50 nM THZ1 + 20uM nutlin-3. N Quantitation of colony formation ability in M. Statistically significant (P < 0.05) are *, (P < 0.01) are **