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. 2001 Jan 8;152(1):111–126. doi: 10.1083/jcb.152.1.111

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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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Localization of endogenous Rho GTPases and RhoGDI. (a) Indirect immunofluorescence: untransfected MDCK cells were grown on coverslips, fixed with 3% paraformaldehyde (i–iv, and vi) or ice-cold methanol/acetone (1:1; v), permeabilized with Triton X-100 (i, ii, and iv) or saponin (iii and vi), and stained for the indicated protein as described in Materials and Methods. (b) Subcellular fractionation: COS-1 cell cytosolic and membrane fractions were analyzed by immunoblot for the indicated proteins, and relative amounts were quantitated by a PhosphorImager. Results are expressed as the percentage of the endogenous protein detected in the membrane fraction and are given as mean ± SEM (n = 8). Bars, 10 μm.