Cell lines and reagents

Human prostate cancer cell lines (LNCaP, VCaP, CWR22Rv1 (referred to as 22Rv1), PC3, and Du145) were purchased from American Type Culture Collection (Manassas, VA, USA). RPMI 1640 media, phenol red-free RPMI 1640 media, Fetal Bovine Serum (FBS), Charcoal Stripped FBS One Shot, 1X phosphate buffered saline (PBS) and 100X Antibiotic-Antimycotic (A/A) solutions were purchased from Gibco-Invitrogen Corporation (Carlsbad, CA). Quick Start Bradford 1X dye reagent, 4X Laemmli sample buffer, a Mini Trans-Blot wet transfer system PVDF membranes and non-fat dry milk blocking grade blocker were purchased from Bio-Rad Laboratories (Hercules, CA). cOmplete, EDTA-free Protease Inhibitor Cocktail Tablets were purchased from Roche Diagnostics (Indianapolis, IN). PARP (9542), pAKT (2920), AKT (4691), pS6 (4858) S6 (2317) primary antibodies and secondary anti-mouse IgG (7076) and anti-rabbit (7074) horseradish peroxidase (HRP)-conjugated antibodies were purchased from Cell Signaling Technology, Inc. (Danvers, Massachusetts). Western Bright ECL western blotting detection kit was purchased from Advansta (Menlo Park, CA).