Bnip 3 sc 56167

Tissues were homogenized in lysis buffer, and proteins (50 μg) were loaded and electroblotted. The blots were probed with polyclonal primary antibodies against anti-Ambra1 (ab69501, Abcam, Cambridge, MA, USA), anti-Beclin-1 (sc-48381), p62 (sc-48373), Pink1 (sc-33796), Parkin (sc-30130), BNIP-3 (sc-56167) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-LC3B (#2775), p53 (#9282), and pAkt (#9271) (Cell Signaling Technology, Danvers, MA, USA) at 4 °C overnight. The primary antibody was visualized by adding a secondary antibody and performing an electroluminescence assay (Amersham Pharmacia Biotech, Piscataway, NJ, USA).

RORα (GTX100029, 1:1000), SQSTM1/p62 (GTX629890, 1:1000), PINK1 (GTX107851, 1:1000), LC3B (GTX127375, 1:1000), Ambra1 (GTX55507, 1:1000), and HIF1α (GTX628480) were from Genetex. VDAC1 (#4661, 1:000), HA (#3724, 1:1000), LC3B (#83506, 1:500), PDH (#3205, 1:1000), ANT2/SLC25A5 (#14671, 1:1000), and HSP60 (#12165, 1:1000) were from Cell Signaling Technology. GAPDH (MAB374 clone 6C5, 1: 10,000, Millipore), Cav-3 (sc-5310, 1:1000), Parkin (sc-32282, 1:1000), and Bnip3 (sc-56167, 1:1000) were from Santa Cruz Biotechnology. Polyclonal goat anti-rabbit IgG-HRP (A9169, 1:5000), polyclonal rabbit anti-mouse IgG-HRP (A9044, 1:5000), and polyclonal rabbit anti-goat IgG-HRP (A5420, 1:5000) were from Sigma-Aldrich.