Hedgehog components and PDGFRα localize to OSE primary cilia. IFM analysis of serum-starved wt OSE cells using antibodies against the Hh components GLI2, SMO, and PTCH1 (A), or PDGFRα (D). Primary cilia (arrows) were detected with anti-acetylated α-tubulin (Acet.tub) antibody. B) WB analysis of wt and cancer OSE cells grown in the presence (0 hour) and or absence (72 hours) of serum. Blots were probed with antibodies against the full-length activator form of GLI2, GLI2(FL), or the C-terminally processed repressor form of GLI2, GLI(R). α-tubulin (α-tub) was applied as loading control. C) RT-PCR showing the expression level of the Hh responsive genes PTCH1 and GLI1 in cultures of wt and cancer OSE cells serum depleted for 72 hours. GAPDH was applied as an internal control. E) WB analysis of wt and cancer OSE cells grown in the presence (0 hour) or absence (72 hours) of serum. The PDGFRα antibody used recognizes two protein bands; #1 is the fully glycosylated form and #2 is the partly glycosylated form of the receptor. α-tub was applied as loading control. IFM, immunofluorescence microscopy; Hh, hedgehog; OSE, ovarian surface epithelium; WB, western blot; wt, wild type.