Anti androgen receptor

Prostates were dissected and sectioned for histological analyses as previously described 11 (link), 36 (link). Hematoxylin and Eosin staining, immunohistochemical analyses, and in situ hybridization were performed on 5-µm thick sections mounted on Superfrost/Plus slides (Fisher Scientific, Pittsburgh, PA). Antigens were retrieved by incubation in citrate buffer (10 mmol/L) for 20 minutes at 100°C or as suggested by antibody manufacturers. The sources and concentrations of primary antibodies used are: anti-α-smooth muscle actin (1:1) from Sigma (St Louis, MO); anti-Vimentin (1:200), anti-E-cadherin (1:200) from Cell Signaling Technology; anti-androgen receptor (1:200) from Santa Cruz; anti-CD31 (1:200) from Abcam; anti-Ki67 (1:500) from Novus Biologicals. For immunofluorescence, the specifically bound antibodies were detected with FITC-conjugated secondary antibodies and visualized under a Zeiss LSM 510 Confocal Microscope. For immunohistochemical staining, specifically bound antibodies were detected with biotinylated anti-Rabbit IgG or biotinylated anti-mouse IgG antibodies (Vector labs). The signal was enhanced using the VECTASTAIN ABC system and visualized with a VECTOR NovaRED Substrate kit. Prostate lesion grading was performed as described 37 (link), 38 (link).