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Bid releases cytochrome c from mitochondria in permeabilized HeLa cells without disruption of membrane potential or protein import. Permeabilized HeLa cells (2 × 105) were incubated in 100 μl of import buffer with the indicated amounts of Bid for 30 min at 22°C. An aliquot was spun down to analyze cytochrome c content, and the remaining sample was used to assess import competence. Cell pellets were extracted with NP-40 lysis buffer and analyzed as described in Fig. 7.
