Nature research paper: Structural basis of mitochondrial membrane bending by the I–II–III2–IV2 supercomplex
, and cell pellets were resuspended in homogenization buffer and lysed in a Dounce homogenizer on ice. Intact mitochondria were isolated first by differential centrifugation of the lysate; following membrane solubilization, the lysate was cleared at 30,000for 30 min at 4 °C, and finally, on a discontinuous sucrose gradient with 15%, 23%, 32% and 60% w/v sucrose in buffer SEM at 14,1371for 60 min at 4 °C in an SW28 rotor.
. In brief, each sample lane from CN-PAGE was incubated with an aqueous solution to reveal protein bands or the presence of active CI and 0.1 mg ml
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