Western Blotting Protein Analysis Protocol

Western blotting was performed as previously described (Liu et al., 2018 (link)). Briefly, cells were lysed in Laemmli sample buffer (cat# 161–0737, Bio-Rad) before SDS–PAGE. Primary antibodies used for western blotting were as follows: anti-FUNDC1 (home-made) (Liu et al., 2012a (link)), anti-LC3 (cat# L8918, Sigma), anti-TIMM23 (cat# 611222, Biosciences), anti-TOMM20 (cat# 612278, Biosciences), anti-STAT3 (cat# 9139, Cell Signaling Technology), anti-phosphorylated STAT3 (cat# 9134, Cell Signaling Technology), anti-STAT5 (cat# 94205, Cell Signaling Technology), anti-phosphorylated STAT5 (cat# 4322, Cell Signaling Technology), anti-HIF2α (cat# NB100-122, Novus Biologicals), anti-βACTIN (cat# 8H10D10, Cell Signaling Technology), and anti-αTUBULIN (cat# ab52866, Abcam). Horseradish peroxidase-conjugated secondary antibodies were used, and signals were detected using an ECL kit (cat# WP20005, Invitrogen).