qPCR analysis of the expression of AR (A), testosterone induced gene Pem (B) and testosterone repressed gene Igfbp3 (B). In A, infection of TM3 cells with retorvirus expressing AR cDNA is shown to lead to AR expression at a level similar to that of LNCaP prostate cancer cells. Significantly different from TM3 cells at p < 0.001 (***). In B, cells were untreated (DMSO vehicle alone, -), or were treated for 24 hr with testosterone (T, 10 nM), MAA (M, 5 mM), or with T + MAA, as indicated. Rhox5 is a testosterone-inducible Sertoli cell marker gene that is also expressed at a low level in Leydig cells in vivo . Igfbp3 is repressed by testosterone, and by MAA , with the latter response also seen in the TM3 cells deficient in AR. Data are mean ± SD values based on n = 3 replicates, with the untreated TM3 cell value set to 1.0. Significantly different from untreated control at p < 0.05 (*) or at p < 0.001 (***); significantly different from T at p < 0.05 (#) or at p < 0.001 (###); and significantly different from MAA at p < 0.001 (+++).