Fig. 7.

AR overexpression inhibits USP14 silencing-induced G₁–S arrest and apoptosis. a Fluorescence-activated cell sorting analysis (FACS) was performed on the indicated breast cancer cells treated with the AR siRNA or control siRNA for 48 h. Cells in each population were calculated from three independent replicates. Mean ± S.D. (n = 3). b The indicated breast cancer cells treated with AR siRNA or control siRNA for 72 h. Flow cytometry analysis with annexinV-FITC/PI staining was used to calculate the apoptotic cells. Apoptotic populations from three independent replicates are shown. Mean ± S.D. (n = 3). *P < 0.05 vs. each vehicle control. c and d Protein lysates were collected from the indicated breast cancer cells exposed to AR siRNA or control siRNA for 48 h. Western blot assay was used to detect the expression of AR, CDK4, CDK2, cyclin D1, PARP, and Bcl-2. e MDA-MB-453 cells stably expressing USP14 or control shRNA were transduced with AR or control vector for 48 h and subjected to FACS analysis. Cells in each population were calculated from three independent replicates. Mean ± S.D. (n = 3). f Protein lysates were collected from MDA-MB-453 cells stably expressing USP14 or control shRNA transduced with AR or control vector for 48 h. Western blot assay was used to detect the expression of CDK4, CDK2, cyclin D1, USP14, and HA-tag. g MDA-MB-453 cells exposed to USP14 siRNA or control siRNA with or without AR or control vector for 72 h were collected. Flow cytometry analysis with annexinV-FITC/PI staining was used to calculate the apoptotic cells. Apoptotic populations from three independent replicates are shown. Mean ± S.D. (n = 3). *P < 0.05 vs. vehicle control. ^#P < 0.05 vs. si-USP14. h Western blot assay was used to detect the expression of PARP, Bcl-2, USP14, and HA-tag. GAPDH was used as an internal control. i MDA-MB-453 cells were exposed to USP14 siRNA or control siRNA with or without AR or control vector for 72 h. j MDA-MB-453 cells were exposed to IU1 with or without AR or control vector for 72 h. MTS assay was used to detect cell viability. Error bars correspond to 95% CI of three independent replicates. *P < 0.05 vs. each vehicle control. ^#P < 0.05 vs. si-USP14 or IU1 treatment