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. 2012 Apr 15;23(8):1593–1604. doi: 10.1091/mbc.E11-11-0900

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© 2012 Oinuma 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 6: — Lipid raft enables efficient binding of Rnd1 to p190 RhoGAP. (A) COS-7 cells were transfected with an expression vector encoding GFP-tagged p190 RhoGAP, and the cell lysate was incubated with GST-fused Rnd proteins. GST proteins used in the pulldown assay were verified by CBB staining. (C, E) Lysates from transiently transfected COS-7 cells with or without MβCD treatment (20 mM, 20 min) were immunoprecipitated with an antibody against Myc. Bound and total proteins were analyzed by immunoblotting with the indicated antibodies. (B, D, F) Relative binding was determined by the amount of bound p190 RhoGAP normalized to the amount of p190 RhoGAP in cell lysates. The results are the mean ± SEM of three independent experiments. **p < 0.01, *p < 0.05 (one-way ANOVA).