Ab192306

BCPAP cells or C643 cells were harvested and lysed with RIPA buffer (Millipore). Immunoblotting was performed in a standard fashion. The following antibodies were used: β-actin (1 : 1000; cat #AF5003; Beyotime, China), NDUFB3 (1 : 500; cat #SC-393351; Santa Cruz, CA), MT-ND1 (1 : 1000, cat #ab181848; Abcam), MT-ND2 (1 : 1000, cat #PA5-37185; Invitrogen), MT-ND3 (1 : 1000, cat #ab192306; Abcam), MT-ND4 (1 : 1000, cat #ab219822; Abcam), MT-ND4L (1 : 1000, cat #PA5-103953; Invitrogen), MT-ND5 (1 : 1000, cat #ab230509; Abcam), MT-ND6 (1 : 1000, cat #PA5-109993; Invitrogen), GPX1 (1 : 1000, cat #ab22604; Abcam), MnSOD (1 : 1000, cat #ab68155; Abcam), CuMnSOD (1 : 1000, cat #ab13498; Abcam), PRDX1 (1 : 1000, cat #NBP1-82558; Novus), PRDX3 (1 : 1000, cat #NBP2-67043; Novus), PRDX6 (1 : 1000, cat #H00009588-M01; Novus), and α-tubulin (1 : 1000; cat# AF0001; Beyotime, China).

The protein extraction and Western blot analysis were performed as previous described.²² The antibody information was as following: kindlin‐2 antibody (13562s, Cell Signaling Technology), Bax antibody (14796, Cell Signaling Technology), Bcl‐2 antibody (ab194583, Abcam), GRP78 antibody (GRP78, 3183, Cell Signaling Technology), CHOP antibody (2895, Cell Signaling Technology), PDI antibody (3501, Cell Signaling Technology), Ero1‐Lα antibody (ab81959, Abcam), Drp‐1 antibody (ab184247, Abcam), Tfam antibody (ab131607, Abcam), ND3 (ab192306, Abcam), Mfn‐2 (9482, Cell Signaling Technology), Fis‐1 (ab71498, Abcam), ATPB (ab170947, Abcam), β actin Antibody (HRP‐60008, Proteintech), Goat anti‐rabbit IgG antibody (SA00001‐2, Proteintech) and Goat anti‐mouse IgG antibody (SA00001‐1, Proteintech). The Image J software was used for quantitative analysing, and relative protein levels were expressed as the intensity ratio of target protein and β actin.

Primary hepatocytes, liver and intestine tissues were collected and lysed in RIPA buffer containing PMSF and phosphatase inhibitor cocktail for 10 min on ice. After determining protein concentrations, we boiled the lysates and subjected them to SDS-PAGE. Subsequently, proteins were transferred onto polyvinylidene fluoride membranes, which were blocked with 5% defatted milk for 1 h at room temperature. The membrane was incubated overnight at 4 °C with primary antibodies and then incubated with secondary antibodies for 1 h at room temperature. Immunoreactive signals were detected with ECL reagent (Thermo Scientific). Image J was used to quantify the Western bands.
Antibodies against SCD1 (2794), FABP1 (13368), CS (14309), COX IV (11967), AMPKα (5832), p-AMPKα (Thr172, 2535), ACC (3662), p-ACC (Ser79, 3661), GAPDH (5174), β-actin (3700), HSP90 (4877), anti-mouse (7076) and anti-rabbit (7074) secondary antibodies were obtained from Cell Signaling Technology. Antibodies to CPT1A (ab128568), ACADL (ab196655), FADS1 (ab126706), mt-ND3 (ab192306), CD36 (ab133625) and GRIM19 (ab110240) were purchased from Abcam. Antibody against Ndufs4 (WH0004724M1) was from Sigma. Antibody to mt-ND2 (LS-C498022) was acquired from Lifespan.