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. 1999 Nov 15;147(4):809–822. doi: 10.1083/jcb.147.4.809

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© 1999 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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PEF is large and proteinaceous. Mitochondria were incubated with tBid (170 ng/ml) in the presence of either buffer B, ultra- or micro-filtered cytosol (10-, 50-, 300-kD, and 0.22-μm filters) normal cytosol, cytosol diluted with buffer B, proteinase K–treated cytosol, or dialyzed cytosol. The control (first sample) did not contain tBid. Ac-DEVD-CHO (10 μM) was added to all samples to maximize PEF activity (see Fig. 9). At the indicated times, aliquots were either pelleted and analyzed for mitochondrial cytochrome c content (bottom) or tested for complex IV accessibility (top).