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. 2001 May 28;153(5):1061–1070. doi: 10.1083/jcb.153.5.1061

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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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CPY* aggregates in the Δjem1 Δscj1 and the kar2-159 mutants. Wild-type (KYSC1, wt), Δjem1 Δscj1 (KYSC4), and kar2-159 (KYSC13) cells were grown to early logarithmic phase at 23°C and then shifted to 37°C for 60 min. Cell extracts were prepared and subjected to sucrose density gradient centrifugation as described in the Materials and Methods. Fractions were collected and proteins were analyzed by SDS-PAGE followed by immunoblotting using an anti-CPY antiserum. Relative amounts of CPY* were quantified. Numbers indicate fractions (from bottom to top) and P represents the pellet fraction recovered in the bottom of the tube. Vertical arrowheads show the positions of apoferritin (440 kD), alcohol dehydrogenase (150 kD), bovine serum albumin (70 kD), and carbonic anhydrase (30 kD).