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. 2001 Mar 5;152(5):877–894. doi: 10.1083/jcb.152.5.877

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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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Golgi elements interact with vimentin filaments in vitro. (A) Recombinant vimentin alone (Vimentin only) or recombinant vimentin incubated with isolated Golgi elements (Vimentin + Golgi) were loaded at the bottom of a sucrose density gradient and subjected to equilibrium centrifugation. An equivalent amount of each fraction was processed by SDS-PAGE, transferred to NC, and immunoblotted with antivimentin and anti-GM130 antibodies. Vimentin remains in the load when centrifuged alone but is recovered in a lower density fraction containing the Golgi marker GM130 when incubated with Golgi elements. (B) Fractions 3 and 4 from A were combined, loaded at the bottom of a sucrose density gradient, and subjected to another round of equilibrium centrifugation. Fractions were collected and analysed as in A. Vimentin remains associated with Golgi elements.