Fig. 5
SUMOylation upregulates AR-dependent transcriptional activity and mesenchymal gene expression in BCa. a Characterizing the SUMO-mimetic AR construct by western blot; asterisk represents AR-SUMO conjugates. b MCF-7 cells were hormone-deprived for 48 h then transfected with probasin and renilla luciferase together with either ARwt or SUMO-fused AR (S-AR). The next day, cells were either untreated or treated with 10 μM Enz, 10 nM R1881 or a combination of both for additional 24 h then harvested and assayed for luciferase activity. Graph B represents fold changes in firefly/renilla ratios for at least three independent experiments, each performed in triplicates and asterisks denote significant changes using One-way ANOVA followed by Tukey’s multiple comparison test. c RNA from cells expressing ARwt or S-AR were used for real-time PCR to assess the expression of mesenchymal, epithelial markers and EMT inducers (ZEB1 and ZEB2). Graph represents fold-changes in indicated mRNA (2ΔΔCt values) for 3 independent experiments; asterisks indicate statistically significant differences in raw ΔCt values when compared to the ARwt + vehicle group using Student’s t-test