Protein extraction and western blot analysis were conducted as described before.31 The primary antibodies included: anti‐TFAM antibody (ab131607, 1:1000), anti‐ATPB antibody (ab14730, 1:5000), anti‐GPX4 antibody (ab125066, 1:3000) and anti‐HMGB1 antibody (ab79823, 1:10000) from Abcam (Cambridge, UK), anti‐BCL2 antibody (3498, 1:1000), anti‐LC3B antibody (3868, 1:1000) and anti‐P62 antibody (5114, 1:1000) from Cell Signaling Technology (Danvers, MA, USA), HRP‐conjugated mouse anti‐ACTB (KC‐5A08, 1:5000), and HRP‐conjugated mouse anti‐GAPDH (KC‐5G5, 1:5000) from Aksomics (Shanghai, China).
The HRP‐conjugated secondary antibodies involved anti‐mouse IgG (KC‐MM‐035, 1:5000) and anti‐rabbit IgG (KC‐RB‐035, 1:5000) from Aksomics (Shanghai, China). The protein bands were visualized with the ChemiDoc MP system (Bio‐Rad, CA). ImageJ software was applied to quantify the band intensity, and the results are presented as the ratio of the target protein to GAPDH or ATCB.
The HRP‐conjugated secondary antibodies involved anti‐mouse IgG (KC‐MM‐035, 1:5000) and anti‐rabbit IgG (KC‐RB‐035, 1:5000) from Aksomics (Shanghai, China). The protein bands were visualized with the ChemiDoc MP system (Bio‐Rad, CA). ImageJ software was applied to quantify the band intensity, and the results are presented as the ratio of the target protein to GAPDH or ATCB.