(A) Cells were lysed in the presence of protease inhibitors, as indicated under Materials and methods. Equal amounts of protein (80 µg) were separated by 10% SDS-PAGE, transferred onto nitrocellulose filters, and blotted with anti-IGF1R, anti-phospho (p)-IGF1R, anti-AR, and anti-tubulin. (B) Effect of 5-Aza treatment on IGF1R promoter activity. P69 and M12 cells were treated with 5-Aza (or left untreated) and, after 24 h, cells were transiently transfected with a proximal IGF1R promoter-luciferase reporter construct [p(−476/+640)LUC], along with a β-galactosidase plasmid. Cells were harvested 48 h after transfection for luciferase and β-galactosidase assays. * p<0.02 versus respective control.