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. 2000 Nov 27;151(5):973–984. doi: 10.1083/jcb.151.5.973

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© 2000 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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Coimmunoprecipitation of HA-Pma1p with Lst1p and Sec24p. Vesicles were generated from lst1-null, HA-Pma1p containing SICs using either a mixture of 10 μg/ml each of Sec23/Lst1p and Sec23/24p (lanes 1) or 20 μg/ml Sec23/24p only (lanes 2). In lanes 3, a mixture of 10 μg/ml each of Sec23/Lst1p and Sec23/24p was used as in lanes 1, but wild-type SICs membranes, not expressing HA-tagged Pma1p, were used. Lanes 4 contained a mixture of the vesicles generated in reactions 2 and 3. After cross-linking, reactions were immunoprecipitated with anti-Lst1p or anti-Sec24p as indicated, and subjected to SDS-PAGE and immunoblot analysis using anti-HA antibodies. Input lanes represent 1% of the cross-linked reactions before immunoprecipitation.