Fig. 6

Synergistic growth inhibition by the co-treatment of enzalutamide and USP14 inhibition involves mechanisms more than suppressing AR-PI3K/AKT signaling. MDA-MB453 and MCF-7 cells exposed to MK2206 (MK, 5 μM), enzalutamide+IU1/USP14 siRNA (I + E or si − +E), the combination of the three agents (MK + I + E or MK + si − +E), or vehicle control DMSO (DM) for 48 h were used for the following assays. a Cell viability assay.*p < 0.05, ^#p < 0.01 vs. other treatment group. b Colony formation assay performed as described in Fig. 2. Representative images are shown. c Representative images of Western blot analysis for the indicated proteins. GAPDH was probed as a loading control. d Prevalence of cell death detected with the same flow cytometry analysis as described in Fig. 2. *p < 0.05, ^#p < 0.01 vs. other treatment group. e representative images of western blot analyses for PARP and cleaved caspase-3 in the total protein extracted from the treated cells