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. 2001 Jan 22;152(2):401–410. doi: 10.1083/jcb.152.2.401

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© 2001 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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Mitochondrial membrane vesicles containing Mmm1p are located in a fraction intermediate in density to outer and inner membrane vesicles. Mitochondria were isolated from YSB107 (Burgess et al. 1994), which expresses the Mmm1p-HA fusion protein. Mitochondria were sonicated and membrane vesicles were separated on sucrose gradients (Pon et al. 1989). Gradient fractions were collected and analyzed by Western blotting using antibodies to Om45p, an outer membrane protein, the β-subunit of the F1-ATPase (F1β), an inner membrane protein, and HA (Mmm1p-HA). Lane 1 represents the top of the gradient.