Figure 2.

Expression of AR isoforms in PCA cells. (A) The relative expression levels of AR, AR3, AR4 and AR5 were quantified using real-time PCR(left panel). The AR level in LNCaP was arbitrarily set as 1. AR3 expression in LNCaP and C-81 were further plotted with a higher resolution. *p<0.05(middle panel). Their expression in two pairs of CW22R xenograft tumors derived from the intact and castrated male mice were also quantified(right panel). *p<0.05. (B) Transcriptional activity of AR isoforms. COS-1 were transfected with ARR2-luciferase reporter together with the indicated expression vector. At 24h-posttransfection, the luciferase activity was measured. Cell lysates were blotted with anti-AR and anti-Tubulin, respectively (bottom). (C) COS-1 were transfected with ARR2-Luciferase reporter along with increasing doses of AR3 or AR expressing vector. At 24h-posttransfection, cells were treated with or without 10nM DHT for 24h and luciferase activities were measured. Cell lysates were blotted with anti-AR and anti-Tubulin, respectively (right panel). (D) LNCaP were infected with (+) or without(−) the lentivirus encoding ARshRNA (shAR) as described previously (26). At 6h-postinfection, cells were transfected with ARR2-Luciferase reporter along with AR3 or the codon-switched wild-type AR(ARcs). At 24h-posttransfection, cells were treated with DHT and casodex(CAS) as indicated for 24h before luciferase activity was measured (left panel). Cell lysates were blotted with anti-AR and anti-Tubulin (Suppl. Fig. 15). Right panel, LNCaP were transfected with AR3 vector or control. At 24h-posttransfection, cells were treated with or without Casodex (+CAS or −CAS), the relative PSA levels were quantified using real-time PCR. The PSA level in the control LNCaP was arbitrarily set as 1. *p<0.05.