Fig. 5

USP36 promotes breast cancer progression via ERα. A Immunoblot analysis showing the expression level of ERα and USP36 in MCF-7 cells stably transduced with shUSP36, transfected with Flag or Flag-ERα plasmid. β-Actin was used as the internal control. B GREB1, PKIB, PS2 and PDZK1 mRNA levels were determined by real time PCR after treatment with Flag or Flag-ERα plasmid for 48 h in MCF-7 cells stably transduced with shUSP36. C MCF-7 cells stably transduced with shUSP36 were transfected with Flag or Flag-ERα plasmid for 48 h. The CCK-8 assays were used to detect the cell viability at the indicated time points. D-E MCF-7 cells stably transduced with shUSP36 were transfected with Flag or Flag-ERα plasmid for 48 h. Transwell assays were used to detect migration ability. E shows the quantitative analysis of the Transwell assays results. F-G MCF-7 cells stably transduced with shUSP36 were transfected with Flag or Flag-ERα plasmid for 48 h. Wound healing assays were used to detect migration ability. G shows the quantitative analysis. H-I MCF-7 cells stably transduced with shUSP36 were transfected with Flag or Flag-ERα plasmid for 48 h. The percentage of apoptotic cells was determined by FACS analysis. PI and Annexin V staining were performed on the cells. I shows the quantitative analysis. Three independent experiments were conducted to get the results shown in Panels A-I. All the data are presented as the means ± SDs. **P < 0.01; ***P < 0.001 for comparisons (Student’s t test)