Fig. 2

USP36 depletion inhibits ERα positive breast cancer progression in vivo and in vitro. A-B Real-time PCR was performed to determine USP36 mRNA levels in MCF-7 and T47D cells following USP36 siRNA treatment 48 h. C-D MCF-7 and T47D cells transfected with siControl or siUSP36 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 scrambled siRNA or two independent USP36 siRNAs for 48 h. F and H show the quantitative analysis of the colony formation assay results. I-L MCF-7 and T47D cells 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 were tested for their migration ability using wound healing assays. N and P show the quantitative analysis. Q-T The percentage of apoptotic cells was determined by FACS analysis after MCF-7 and T47D cells were treated with USP36 siRNA for 48 h. PI and Annexin V staining were performed on the cells. U A representative image of a tumor derived from a nude mouse injected with stably transfected shControl or shUSP36 MCF-7 cells is shown. V-W The tumor volume (V) and weight (W) in nude mice subcutaneously inoculated with stably transfected shControl or shUSP36 MCF-7 cells.Three independent experiments were conducted to obtain the results shown in Panels A-W. All the data are presented as the means ± SDs. *P < 0.05; **P < 0.01; ***P < 0.001 for comparisons (Student’s t test)