Overexpression of Hedgehog signaling members, including Patch1, Smo, Gli1, Gli2, and Gli3, in pCX-shh-IG-injected prostates at day 90 after injection. The lower pictures are magnifications of the boxed areas shown in the upper pictures. Hedgehog signalling members were expressed in the PIN (F-J) and CaP (K-O) of the pCX-shh-IG-injected prostates, in contrast to the absence of evident signal in the pCX-IG-injected vehicle controls (A-E; basal/stem cells indicated by arrows). Note that some cells positive for Hedgehog signalling (indicated by arrowheads in F-J) appeared to be basal/stem cells in morphology. Western blot analysis (P) confirmed that Hedgehog signalling members were highly expressed in the pCX-shh-IG- injected prostates, as compared to those sham injections with 0.9% saline or vehicle controls with pCX-IG vector. Tubulin detections served as loading controls. Scale bars: 10 μm in lower E, lower J and lower O. CaP: prostate cancer; PIN: prostatic intraepithelial neoplasia; Gli2-act: active form of Gli2; Gli2-rep: repressed form of Gli2; Gli3-act: active form of Gli3.