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. 2012 Jul 15;23(14):2793–2804. doi: 10.1091/mbc.E12-02-0103

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© 2012 Iwasawa et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 7: — Afadin is localized to the membrane by active R-Ras in a RA domain–dependent manner. (A) Neuro2a cells were transfected with GFP-tagged afadin FL or ΔRA mutant together with HA-tagged R-Ras QL, and the cytosol (C) and the membrane (M) fractions were analyzed by immunoblotting. Na⁺/K⁺-ATPase and ERK were used as markers of membrane and cytosol, respectively. Expression levels of the constructs were verified by immunoblotting of the cell lysates (right). (B) The membrane/cytosol ratio of GFP-tagged afadin was analyzed. Data are presented as the means ± SEM of three independent experiments (*p < 0.05, one-way ANOVA with Dunnett's post hoc test).