Figure 4

Gli overexpression affects androgen regulated gene expression in androgen-deprived LNCaP cells. (A) RNAs from control (Vec) or Gli1 or Gli2 (Gli2ΔN) overexpressing LNCaP cells cultured in androgen-depleted medium for 72 hrs were assayed by real-time qPCR for expression of Gli1, Gli2, KLK2 and KLK3. Bars represent the means of three experiments ± S.E. (* = P < 0.05 compared to vector control). (B) Cells were infected with a Gli or Probasin (PRB) reporter with CMV-GFP and switched to androgen-depleted medium for 72 hrs. Cell extracts were quantified for luciferase that was normalized by GFP intensity. Bars represent the means of triplicate experiments ± S.E. (* = P < 0.05 compared to vector control). (C) Western blot shows that Gli1 or Gli2 overexpression does not affect expression of AR protein. (D) Gli overexpression enables androgen independent cell growth. Control (Vec) or Gli1 or Gli2 (Gli2ΔN) overexpressing LNCaP cells were cultured in androgen depleted medium for 12 days and growth was measured by WST-1 assay and compared to Day 0. Bars represent the means of three experiments ± S.E. (* = P < 0.05 compared to vector control).