Anti tom70

Anti-β-Catenin (catalog no. 610153) and anti-CD133 antibody (catalog no. 566598) was purchased from BD Bioscience. Anti-PGK1 (catalog no. 68540 S), anti-EEA1 (catalog no. 3288 S), anti-β-actin (catalog no. 4970), anti-H3 (catalog no. 4499) and anti-H3K4me3 (catalog no. 9751 S) antibodies were from Cell Signaling Technology. Anti-ZIC2 (catalog no. ARP35821_P050) antibody was purchased from Aviva Systems Biology. Anti-TOM40 (catalog no. 18409-1-AP), anti-TOM70 (catalog no. 14528-1-AP), anti-c-MYC (catalog no. 10828-1-AP) and anti-AXIN2 (catalog no. 20540-1-AP) antibodies were from Proteintech Group, Inc. Goat anti-Mouse IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 594 antibody (catalog no. A-11005), Goat anti-Rabbit IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 488 (catalog no. A-11008), Goat anti-Rabbit IgG (H + L) Cross-Adsorbed Secondary Antibody, Alexa Fluor™ 647 (catalog no. A-21244) were purchased from Invitrogen. HRP-conjugated Affinipure Goat Anti-Mouse IgG(H + L) antibody (catalog no. SA00001-2) and HRP-conjugated Affinipure Goat Anti-Rabbit IgG(H + L) antibody (catalog no. SA00001-2) were purchased from Proteintech Group, Inc. Polymer HRP and AP detection kits were from Beyotime Biotechnology. Biotin labeled RNA mix (catalog no. 11685597910) was from Roche.

Blue native polyacrylamide gel electrophoresis (BN-PAGE) was conducted as mentioned before [27 (link)]. For mitochondrial supercomplexes, 1% digitonin (Sigma-Aldrich) was used to dissolve the membrane protein from patient and controls, then 3.5–16% gradients polyacrylamide gel to separate the supercomplexes. For mitochondrial complexes, 1% dodecyl maltoside (DDM, Sigma-Aldrich) was used and then 3–11% gradient polyacrylamide gel to separate the complexes. The proteins were transferred on the PVDF membrane (Bio-Rad, USA), blocked with 5% no-fatty acid milk (Mengniu, China), incubated with the first and second antibodies, and then detected the protein signals using. The following antibodies were used: anti-Grim19 (1:1000, Sigma-Aldrich), anti-SDHA (1:3000, Sigma-Aldrich), anti-UQCRC2 (1:2000, Sigma-Aldrich), anti-MT-COI (1:2000, Sigma-Aldrich), anti-ATP5A (1:3000, Sigma-Aldrich), anti-NDUFB6 (1:2000, Sigma-Aldrich), anti-NDUFS3 (1:2000, Sigma-Aldrich), anti-TOM70 (1:2000, Proteintech, China), anti-mouse IgG HRP linked (1:2000, Cell Signaling Technology), anti-rabbit IgG HRP linked (1:2000, Cell Signaling Technology), anti-mouse IgG, AP (1:2000, Cell Signaling Technology).