Western Blot Analysis of Protein Markers

30–60μg of protein were separated on precast tris-glycine gels (Invitrogen) and transferred to PVDF membrane. Membranes were blocked with 5% BSA/TBST. Primary antibodies, after incubation in 5% BSA/TBST, were detected using HRP conjugated secondary antibodies in chemiluminescent solution using the Quantity One imaging software on a Bio-Rad Gel Docking system. Primary antibodies: Rabbit mAb Bnip3 (EPR4034) from Abcam, rabbit anti-P-AktSer473, rabbit mAb P-Akt308 (C31E5E), rabbit mAb Akt (pan) (C67E7), mouse mAb Histone H3 (96C10), rabbit mAb P-Ser555 ULK (D1H4, #5869), rabbit mAb ULK (D8H5, #8054), rabbit mAb P-AMPKα (Thr172) (40H9, #2535), and rabbit anti-AMPKα (#2532) from Cell Signaling Technology, rabbit anti-LC3B (NB100–2200) from Novus Biologicals, mouse mAb GAPDH from Millipore, rat anti-integrin α6 (GoH3) and mouse anti-HIF1α from BD Pharmingen, mouse mAb AR (441) from Santa Cruz, mouse anti-α tubulin and β-actin-HRP mouse mAb from Sigma-Aldrich), and rabbit anti-integrin α6 (AA6A, A6NT).^{12 (link), 35 (link)}