Rabbit anti ho 1

Western blots were performed on liver tissue or hepatocytes. Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred onto a nitrocellulose membrane. The membrane was blocked for 1 hour in TBS-Tween 20 with 5% milk, followed by immunostaining with dilutions of primary antibodies [mouse anti-HIF-1a (Novus, Littleton, CO), rabbit anti-HO-2 (Abcam), rabbit anti-HO-1 (Affinity Biosciences, Cincinnati, OH), mouse anti-COX IV (Abcam), mouse anti-beta actin (Abcam), and rabbit anti-total or mouse anti-phospho JNK (Cell Signaling Technology, Danvers, MA)] in 1% milk in phosphate-buffered saline- (PBS-) Tween from 1 hour to overnight at 4°C. Secondary antibodies conjugated with horseradish peroxidase and washed for 1 hour in TBST before being developed with the SuperSignal West Pico chemiluminesence kit (Thermo Fisher Scientific, Waltham, MA). Beta actin was used as a loading control for all whole cell experiments.