Figure 6.

AR is recruited to the GNMT ARE in an androgen-dependent manner. (A) Diagramatic representation of the PSA (top) and GNMT (bottom) promoter regions. AREs are highlighted in boxes and arrows indicate the location of PCR primer pairs. (B) LNCaP cells were cultured in RPMI medium lacking phenol red supplemented with 10% DSS for 72 h followed by addition of 10 nM R1881 or ethanol for 1 h. Cells were then cross-linked, lysed and sonicated. Immunoprecipitations were performed using an antibody specific for AR or rabbit IgG. Immunoprecipitated DNA was reverse cross-linked and recovered by phenol/chloroform extraction. Real-time PCR was carried out using primers for the regions indicated in part A. The PSA enhancer region was chosen as a positive control, and a region distant from ARE elements as a negative control. Results show the mean values obtained from ChIP analysis of two replicates.