Fig. 5

The treatment of combining enzalutamide with and IU1 or USP14 knockdown induces the downregulation of AR and inhibits AR-related signaling pathways. a MDA-MB453 and MCF-7 cells treated with the inidicated agents for 48 h as described in Fig. 3 were harvested for extraction of total proteins which were used for western blot analyses of the indicated proteins. b MDA-MB453 cells were treated with the indicated agents for 24 h as described in Fig. 3 and then fixed and processed for immunostaining for AR (red). Nuclei were counter-stained with DAPI (blue). Fluorescence microscopy was then used to record the expression and distribution of endogenous AR. The images shown are representative of three independent experiments. Scale bar, 50 μm. c-e MCF-7 and MDA-MB453 cells were transfected with luciferase reporter plasmid for 24 h. Then cells were treated with enzalutamide and USP14 inhibitor or siRNA. Protein lysates were collected, followed by dual-luciferase assay for luciferase activity. *p < 0.05, ^#p < 0.01 vs. each treatment alone