Protein extracts were isolated from the hearts of 15-day-olds extracted with RIPA lysis buffer (50 mM Tris-HCl at pH 7.4, 150 mM NaCl, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS, 1 mM dithiothreitol (DTT), 5 mM EDTA, and Halt Protease and Phosphatase Inhibitor Cocktail from Thermo Fisher Scientific), and the protein concentration was measured using DC Protein Assay kits (Bio-Rad).
Equal quantities of proteins were resolved by SDS-PAGE gels and blotted onto polyvinylidene difluoride membranes (Millipore). Then, the membranes were blocked with 2% bovine serum albumin (BSA) before incubation with the primary and secondary antibodies. The immunoblots were visualized by using Luminata -Immobilon Crescendo Western HRP Substrate (Millipore) on a ChemiDoc MP station (Bio-Rad). Image analysis was performed using Image Lab software version 5.2 (Bio-Rad). The antibodies used for the western blots were as follows: anti-Srsf3 (Abcam, ab198291) anti-Gapdh (Cell Signaling, 2118), anti-Tim50 (Santa-Cruz, sc-393678), anti-Tom20 (Abcam, ab186735), Total OXPHOS antibodies (Abcam, ab110413), and anti-Uqcrc1 (ThermoScientific, PA5-21394).
Equal quantities of proteins were resolved by SDS-PAGE gels and blotted onto polyvinylidene difluoride membranes (Millipore). Then, the membranes were blocked with 2% bovine serum albumin (BSA) before incubation with the primary and secondary antibodies. The immunoblots were visualized by using Luminata -Immobilon Crescendo Western HRP Substrate (Millipore) on a ChemiDoc MP station (Bio-Rad). Image analysis was performed using Image Lab software version 5.2 (Bio-Rad). The antibodies used for the western blots were as follows: anti-Srsf3 (Abcam, ab198291) anti-Gapdh (Cell Signaling, 2118), anti-Tim50 (Santa-Cruz, sc-393678), anti-Tom20 (Abcam, ab186735), Total OXPHOS antibodies (Abcam, ab110413), and anti-Uqcrc1 (ThermoScientific, PA5-21394).