Figure 3.
Detection of AR3 in hormone-insensitive PCA cells. (A) COS-1 were transfected with AR3 or AR vector. Total protein lysates were immunoblotted with anti-AR3 and anti-AR, respectively. (B) CWR-R1 and 22Rv1 were infected with lentivirus encoding AR3shRNAs (shAR3-1, −2) or the scrambled control (shAR3-sc). At 48h-postinfection, cell lysates were subjected to immunoblotting with anti-AR and anti-AR3, respectively. COS-1 overexpressing AR3 was used as a positive control (first lane). (C) CWR-R1 lysates were split into three equal aliquots and immnuoprecipitated with anti-AR3, control IgG and anti-AR, respectively. The resultant immunoprecipitates and the input total cell lysates (TCL) were immunoblotted with anti-AR. (D) Total cell lysates of a panel of PCA cells were blotted with anti-AR3 and anti-Actin, respectively (top panel). Bottom panel, extracts of two pairs of CW22R tumor xenografts derived from the intact and castrated male mice were blotted with anti-AR3 and anti-Tubulin, respectively.