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. 2014 Feb 1;25(3):380–396. doi: 10.1091/mbc.E13-07-0363

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© 2014 Georgess 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 9: — RhoE is cytoplasmic and inhibits Rock-I from phosphorylating cofilin. (A) Micrograph of a day-5 BM-OC expressing RhoE-eGFP (green) 24 h posttransfection and stained for F-actin (red), showing ubiquitous, cytoplasmic localization of RhoE. (B) Immunoblotting of total cofilin, phosphorylated cofilin at Ser-3 (p(Ser-3)-cofilin), Rock-I, and β-tubulin from total cell lysate of day-5 BM-OCs that were serum induced for 2 h with or without 10 μM Y-27632 (a Rock inhibitor) after 2 h of serum starvation. Quantification of protein expression of p(Ser-3)-cofilin normalized to total cofilin reveals 1.86-fold increase in RhoE^gt/gt OCs compared with RhoE^+/+ OCs without affecting overall Rock-I levels. Diagrams show mean ± SEM from three independent experiments.