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. 2022 Feb 14;13(2):149. doi: 10.1038/s41419-022-04604-3

Fig. 3. RNF187 depletion increases the P53 protein level and P53 target gene expression in breast cancer cells.

Fig. 3

A, B RNF187 depletion increases the P53 protein level but not the mRNA level in MCF-7 cells. MCF-7 cells were transfected with siControl or siRNF187. After 48 h, the cells were harvested for western blot analysis. RNF187 and P53 protein levels were determined by western blot analysis. Actin was used as the internal control. The P53 mRNA level was determined by qPCR, while 36B4 was used as the internal control. Each group was analyzed in triplicate. *P < 0.05; **P < 0.01; ***P < 0.001 for gene expression comparisons. C RNF187 depletion increases P53 target gene expression. MCF-7 cells were transfected with siControl or siRNF187. After 48 h, total RNA was extracted for gene expression analysis. Each group was analyzed in triplicate. *P < 0.05; **P < 0.01; ***P < 0.001 for target gene expression comparison. D RNF187 depletion increases the P53 protein level in MCF-7 cells under both vehicle and cisplatin treatment conditions. MCF-7 cells were transfected with siControl or siRNF187. After 48 h, the cells were treated with 10 μM cisplatin or vehicle for 6 h. Then, the cells were harvested for western blot analysis. RNF187 and P53 protein levels were determined by western blot analysis. Actin was used as the internal control. E RNF187 depletion increases the P53 protein level in MDA-MB-175 cells under both vehicle and cisplatin treatment conditions. MDA-MB-175 cells were transfected with siControl or siRNF187. After 48 h, the cells were treated with 10 μM cisplatin or vehicle for 6 h. Then, the cells were harvested for western blot analysis. RNF187 and P53 protein levels were determined by western blot analysis. Actin was used as the internal control. F RNF187 depletion increases the P53 protein level in ZR751 cells under both vehicle and cisplatin treatment conditions. ZR751 cells were transfected with siControl or siRNF187. After 48 h, the cells were treated with 10 μM cisplatin or vehicle for 6 h. Then, the cells were harvested for western blot analysis. RNF187 and P53 protein levels were determined by western blot analysis. Actin was used as the internal control. G RNF187 depletion increases P53 target gene expression under both vehicle and cisplatin treatment conditions. MCF-7 cells were transfected with siControl or siRNF187. After 48 h, 10 μM cisplatin was added to treat the cells for 6 h. Total RNA was extracted for gene expression analysis. Each group was analyzed in triplicate. *P < 0.05; **P < 0.01; ***P < 0.001 for target gene expression comparison. H RNF187 depletion increased P53 target gene expression under both vehicle and cisplatin treatment conditions. MDA-MB-175 cells were transfected with siControl or siRNF187. After 48 h, 10 µM cisplatin was added to treat the cells for 6 h. Total RNA was extracted for gene expression analysis. Each group was analyzed in triplicate. *P < 0.05; **P < 0.01; ***P < 0.001 for target gene expression comparison. I RNF187 depletion increased P53 target gene expression under both vehicle and cisplatin treatment conditions. ZR751 cells were transfected with siControl or siRNF187. After 48 h, 10 μM cisplatin was added to treat the cells for 6 h. Total RNA was extracted for gene expression analysis. Each group was analyzed in triplicate. *P < 0.05; **P < 0.01; ***P < 0.001 for target gene expression comparison.