Fig. 2

Transfection control of 93RS2 Sertoli cells. a 24 h after transfection, transfected (a) and non-transfected (b) cells as negative control were fixed for IF experiments. left Incubation with rabbit anti-GFP antibody showed successful transfection of almost 80 % of cells. right No staining signal was detectable in non-transfected cells. Scale bars in main image: 200 µm, detail: 25 µm. DAPI counterstain. b Western Blot analysis revealed AR protein in transfected Sertoli cells at approx. 135 kDa (1) and in human testis tissue at the expected molecular weight of 110 kDa (2). The higher protein weight measured in transfected cells is due to coupling of AR with GFP. c Expression of human AR mRNA was tested in human testis homogenate (1), transfected (2) and non-transfected cells (3). AR mRNA was detected in the positive control and transfected 93RS2hAR17 cells, but not in non-transfected cells and the NTC (lane 4). d To control the CAG repeat length in transfected 93RS2 cells, we performed high-resolution PAGE. Three different passages of 93RShAR17 cells (lanes 1–3) were analysed and revealed a band for human AR at 185 bp by using two different DNA ladders. By sequencing, 185 bp was shown to be typical for the presence of 17 CAG repeats. Lane 4 no template control (NTC)