Fig. 3: FZR1 promoting chemotherapy drug induced apoptosis by regulating the stability of p53.

a RNA seq was performed in control and FZR1 ko T-47D cells treated with cisplatin. Heatmap clustered on expression profiles created based on log2 transformed counts to identify consistent changes in various samples. b GSEA analysis of apoptosis and p53 pathway relevant gene sets enriched among genes up- and down-regulated in control and FZR1 ko T-47D cells treated with cisplatin. c Heatmap clustered on apoptosis and p53 pathway relevant gene sets expression profiles in control and FZR1 ko T-47D cells treated with cisplatin. d The mRNA levels of p53 in control and FZR1 ko T-47D cells with or without cisplatin treatment were measured by qPCR. e Western blot was performed to evaluate the stability of p53 in control and FZR1 ko T-47D cells treated with cisplatin and proteasome inhibitor MG132. f IF of p53 was performed in control and FZR1 ko T-47D cells treated with cisplatin or cisplatin and proteasome inhibitor MG132. A relative quantification of the number of p53 positive cells ± SD of three independent experiments is shown in the histogram on the right. **p < 0.01; scale bar 30 or 10 µm. g The protein stability of p53 was evaluated in control and FZR1 ko T-47D cells treated with cisplatin, proteasome inhibitor MG132 and protein synthesis inhibitor CHX for 5, 15, 30, and 60 min. A representative western blot of three independent experiments is shown. h IP using p53 antibody was performed in control and FZR1 ko T-47D cells expressing ubiquitin with HA tag treated with cisplatin and proteasome inhibitor MG132. The precipitate was detected by western blot using HA antibody.