Fig. 4

Lethality mediated by nutlin-3 + THZ1 required for p53 expression. A si-p53 was transfected into MCF-7 cells previously which would following be given 50 M THZ1and 10uM nutlin-3 treatment for 48 h. Western blot was conducted to display the expression of PARP, cleaved PARP, p53 and b-actin in MCF-7 cell. B Quantity of cleaved PARP and p53 in A. C CCK8 assay was used to measure the cell viability of MCF-7 cell treated with si-p53 or si-control combining 50 M THZ1and 10uM nutlin-3 for 24 h and 48 h. D Colony formation ability measurement of MCF-7 cell after transfecting si-p53 or si-control, then added with 50 nM THZ1 + 10uM nutlin-3. E Quantitation of colony formation ability in D. F Apoptosis was analyzed by flow cytometry after MCF-7 cells that transfected with si-p53 or si-control were treated with 50 nM THZ1 + 10uM nutlin-3 for 48 h. G Quantity of apoptosis in F. H Western blot was utilized to detect PARP, cleaved PARP, p53 and b-actin protein expression in MCF-7 cells which were treated with si-p53 combining 50 nM THZ1 + 0.5uM doxorubicin or 1.25uM LDC4297 + 0.5uM doxorubicin. I Quantity of cleaved PARP and p53 in H. J CCK8 assay was used to measure the cell viability of MCF-7 cells treated with si-p53 combining 50 nM THZ1 + 0.5uM doxorubicin or 1.25uM LDC4297 + 0.5uM doxorubicin for 24 h. K Colony formation ability measurement of MCF-7 cell after transfecting si-p53 or si-control, then added with 50 nM THZ1 + 0.5uM doxorubicin or 1.25uM LDC4297 + 0.5uM doxorubicin. L Quantitation of colony formation ability in K. M Apoptosis was analyzed by flow cytometry after MCF-7 cells that transfected with si-p53 or si-control were followed by 50 nM THZ1 + 0.5uM doxorubicin or 1.25uM LDC4297 + 0.5uM doxorubicin treatment for 24 h. N Quantity of apoptosis in M. Statistically significant (P < 0.05) are *, (P < 0.01) are **