Semidenaturing detergent agarose gel electrophoresis (SDD-AGE) was performed as previously described.14 (link) In brief, crude mitochondria were resuspended in 1× sample buffer (0.5× TBE, 10% glycerol, 2% SDS, and 0.0025% bromophenol blue) and loaded onto a vertical 1.5% agarose gel (Bio-Rad). After electrophoresis in running buffer (1×TBE and 0.01% SDS) for 50 min with a constant voltage of 100 V at 4 °C, the proteins were transferred to an immobile membrane (Millipore) for immunoblot analysis.
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