Figure 5. MYSM1 interacts with AR and inhibits activation of Akt/c-Raf/GSK-3β signaling in prostate cancer. (A) qRT-PCR analyses for MYSM1 and AR mRNA levels following transfection of NC/siMYSM1 into C4-2 and 22Rv1 cells. Data are shown as mean ± SEM of 3 replicates. (B) Western blot analyses for MYSM1 and AR protein levels in C4-2 and 22Rv1 cells treated with shNC/shMYSM1. (C) Representative IHC staining images of AR in xenograft tumor tissues. Scale bars are 30 μm. (D) Co-IP assays of MYSM1 and AR in C4-2 cells with (down) or without (up) siMYSM1 treatment. (E) Correlation of MYSM1 with PHLPP1 (up) and PTEN (down) mRNA levels in prostate cancers. Pearson correlation coefficients and significance levels are indicated. Data were collected from TCGA database and analyzed via LinkedOmics bioinformatics. (F) qRT-PCR analyses for MYSM1 and PHLPP1 mRNA levels following transfection of NC/siMYSM1 into C4-2 cells. Data are shown as mean ± SEM of 3 replicates. (G) Western blot analyses of lysates from C4-2 cells treated with shNC/shMYSM1 were probed with the indicated antibodies. n.s = no significance, * P < 0.05 and *** P < 0.001 (Student’s t-test).