Anti sam50

All antibodies utilized in this study: anti-CHCHD4 (Proteintech, #21090-1-AP); anti-β-Actin (Proteintech, #81115-1-RR); anti-α-SMA (CST, #19245S); anti-citrate synthetase (Abcam, # ab129095); anti-Vimentin (Abcam, #ab92547); anti-PCNA (Santa Cruz, #sc-56); anti-Ki67 (Abcam, #ab15580); anti-Opa1 (Santa Cruz, # sc-393296); anti-Drp1 (Santa Cruz, # sc-271583); anti-Mfn1 (Santa Cruz, #sc-166644); anti-SAM50 (Abcam, #ab246987); anti-DYKDDDDK Tag (CST, #14793); anti-HA (CST, #3724).

Cells were harvested by 0.25% trypsin (Gbico), cell suspension was then centrifuged, pellet cells were resuspended with 1× sample buffer, and boiled for 12 min. The protein samples were separated by SDS-PAGE and transferred into 0.45 μm PVDF membranes, 2 h later, membranes were blocked by 5% non-fat milk for 1 h, then incubated with primary antibody 30 min-2h in 5% non-fat milk followed by incubation with 1:5000 HRP secondary antibodies for 1 h. Detection of immunoreactive protein on Fuji x-ray film using ECL chemiluminescent substrate (Bio-Red). The following primary antibodies were used: anti-GAPDH and anti-Beta-Tubulin were obtained from Santa Cruz Biotechnology; anti-COX4, anti-COX2, anti-ATAD3A, anti-Mic60, anti-Yme1L, and anti-Tom20 were from Proteintech; anti-OPA1 was obtained from BD Biosciences, anti-Sam50 was obtained from Abcam; anti-Mic19 was from Abclonal; anti-Mic10 was from Invitrogen.