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. 2001 Feb 5;152(3):519–530. doi: 10.1083/jcb.152.3.519

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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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Subcellular fractionation of Vps30p, Vps38p, and Vps34p. AKY106 (wild type), AKY110 (Δvps34), AKY111 (Δvps30), AKY112 (Δapg14), and AKY126 (Δvps15) cells, each bearing pKHR65 (VPS38-3xHA), and AKY113 (Δvps38) cells bearing pRS313 (empty vector) were grown in SC without histidine at 28°C. Cell lysates were subjected to subcellular fractionation by differential centrifugation as described in Materials and Methods. Total cell lysate (lane 1), LSP (lane 2), HSP (lane 3), and HSS (lane 4) fractions were subjected to SDS-PAGE, followed by immunoblotting with anti-Vps30p (A), anti-HA (16B12) (B), anti-Vps34p (C), or anti-Pho8p (D) antibodies. Relative amounts of each fraction were indicated. The values in A and C were averages of three independent experiments.