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. 2000 Oct 30;151(3):685–696. doi: 10.1083/jcb.151.3.685

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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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Rac-induced focal complexes contain β3 integrin and vinculin and form by a mechanism that does not involve calpain. BAE cells were grown to 80% confluency and transiently transfected with plasmid encoding HA-tagged constitutively active Q61LRac. Cells were detached and replated on fibronectin-coated coverslips in the presence of calpeptin (100 μg/ml), MDL (250 μM), or corresponding volume of DMSO for 1 h, fixed, and permeabilized. Cells were stained with antibodies against β3 integrin. Transfected cells were detected with anti–HA epitope antibody, actin filaments were detected with TRITC-labeled phalloidin. Bar, 20 μm.