Figure 3. Sox2 is negatively regulated by Androgen Receptor (AR) signaling in prostate epithelial cells (PrECs) and human embryonic stem cells (hESCs).

A) To test whether AR signaling impacted Sox2 expression, we ectopically expressed AR (LV-AR) and control lentivirus (LV-Control) in two different Sox2-positive PrEC lines (#1 and #2). Western blotting shows that expression and further ligand activation of AR using 1 nM R1881 results in decreased Sox2 protein expression and decreased p63 expression. GAPDH was used as a loading control. B) To test whether AR-mediated inhibition of Sox2 expression was specific to PrECs or could also occur in Sox2-positive human ES cells, we treated the WA01(H1) human ES cell line with androgen (1 nM R1881). Western blotting documents detectable endogenous AR expression in hESCs, which increases in response to androgen. β-Actin was used as a loading control. C) AR activation in hESCs results in a significant decrease in Sox2 mRNA expression as measured by qPCR (* indicates p<0.05). This was compared to untreated and vehicle treated cells (EtOH).