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. 1999 Jul 26;146(2):333–344. doi: 10.1083/jcb.146.2.333

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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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The localization of GFP-Sec1p to sites of exocytosis is altered in sec mutants. SEC+ GFP-Sec1p (NY1699), SEC+; No GFP (NY 1700), sec4-8 GFP-Sec1p (NY1697), sec4-8; No GFP (NY1701) and sec18-1 GFP-Sec1p (NY1698) were grown to early log phase. 5 A₆₀₀ units were concentrated and incubated at 37°C for 10 min (or incubated at 25°C), then diluted 10-fold into ice-cold wash buffer. Cells were fixed with methanol and acetone, washed with ice-cold PBS, and visualized by epifluorescence microscopy.