LNCaP cells were seeded onto plates overnight in RPMI-1640 medium containing 10% fetal bovine serum, then switched to an androgen-depleted medium as was previously described [110] containing Vec or GANT-58 or GANT-61 dissolved in dimethyl sulfoxide at the indicated concentrations and was incubated for an additional 72 h. RNAs were then extracted from these cells and were assessed by quantitative real-time PCR for the expression of KLK2 or KLK3 (prostate-specific antigen), as described, and the results are normalized to the expression of GAPDH in the same samples. Each point indicates the means ± standard deviation from triplicate cultures.
GANT: Gli antagonist; KLK: Kallikrein-related peptidase; Ptch: Patched; Vec: Dimethyl sulfoxide vehicle only.