Wnt-11 promotes neuroendocrine-like differentiation in PCa cells. (a) Extracts from control shRNA and WNT11 shRNA LNCaP cells cultured for 3 days in hormone-depleted medium were probed by western blotting for neuron-specific enolase (NSE). The blot was then stripped and re-probed for HSP60. (b) RT-PCR for WNT11, NSE, ASCL1 and GAPDH expression after transfection of LNCaP cells either with empty vector (V) or Wnt-11 expression plasmid. (c) RT-PCR for NSE, ASCL1 and GAPDH expression levels in LNCaP cells transfected either with empty vector (V, lane 1) or Wnt-11 expression plasmid (lanes 2-6) and cultured in the presence of carrier (DMSO, lanes 1 and 2), H89 (lane 3), KT5720 (lane 4) Rp-cAMPs (lane 5) or PKI (lane 6). (d) Immunostaining for Wnt-11 (red) and AR (green) in LNCaP cells transfected with Wnt-11 expression plasmid. Red arrows indicate a cell with high Wnt-11 and low AR. TO-PRO-3 was used to stain nuclei. (e) RT-PCR for WNT11, NSE and GAPDH in PC3 cells transfected with control siRNA (Ct si) or WNT11 siRNA (WNT11 si). (e) RT-PCR for WNT11, NSE and GAPDH in RWPE-1 cells transfected as in (b) with empty vector (V) or Wnt-11 expression plasmid.