Fig. 3

USP36 instead of USP36 C131A overexpression promotes ERα positive breast cancer progression in vivo and in vitro. A-B Immunoblot analysis showing the expression level of USP36 in MCF-7 and T47D cells transfected with Flag or Flag-USP36 WT or Flag-USP36 C131A plasmid. β-Actin was used as the internal control. C-D MCF-7 and T47D cells transfected with Flag or Flag-USP36 WT or Flag-USP36 C131A plasmid for 48 h were tested for viability using the CCK-8 assay at the indicated time points. E–H Colony formation (left panel) of MCF-7 and T47D cells transfected with indicated plasmid for 48 h. F and H show the quantitative analysis of the colony formation assay results. I-L MCF-7 and T47D cells transfected with indicated plasmid for 48 h, were tested for their migration ability using Transwell assays. J and L show the quantitative analysis of the Transwell assays results. M-P MCF-7 and T47D cells transfected with indicated plasmid for 48 h, were tested for their migration ability using wound healing assays. N and P show the quantitative analysis. Q A representative image of a tumor derived from a nude mouse injected with stably MCF-7 cells as indicated is shown. R-S The tumor volume (R) and weight (S) in nude mice subcutaneously inoculated with stably transfected MCF-7 cells as indicated. Three independent experiments were conducted to get the results shown in Panels C-S. All the data are presented as the means ± SDs. *P < 0.05; **P < 0.01; ***P < 0.001 for comparisons (Student’s t test)