Figure 2.

Expression of AR mRNA (a), androgen target gene FKBP5 (b), and AR protein (c) in fibroblasts and LNCaP prostatic cancer cells. The cells were split onto 6-well plates, and after 24 h, the medium was changed to steroid-depleted medium for 6 h. A half of the wells were treated with vehicle (0.1% ethanol) (−) and a half with 1 nM R1881 (+) for 18 h before immunoprecipitation or RNA extraction. Samples XX31B, XY31A, and XX54A are control fibroblasts. The patient-derived fibroblasts are samples T1 and T2. GAPDH served as the reference gene for quantification of AR and FKBP5 mRNA (panels a and b). The expression of all samples was normalized to the control sample XX31B vehicle treatment. The bars represent mean ± SD of 3–5 independent samples. In (c), AR was immunoprecipitated with rabbit polyclonal α-AR¹⁷ and detected in western blotting with mouse monoclonal α-AR 441 recognizing AR amino acids 299–315 in the AR N-terminal domain. α-GAPDH was used to control the loading of input samples. The samples in the different blots are from the same experiment. The blots have been cropped; full-length blots are presented in Supplementary Figure S1.