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. 2017 Jun 15;8:15838. doi: 10.1038/ncomms15838

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Copyright © 2017, The Author(s)

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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(a–c) Cultured mouse MKs were transduced with Raichu-Cdc42 lentiviral vector. F-actin was stained with phalloidin (blue); GPIbβ, red; DAPI, cyan. Cdc42 activation was visualized by colour-coded FRET. High ratio values (red) correlate with higher Cdc42 activity. Active Cdc42 localizes with the polarized F-actin/DMS complex and its activity is decreased in Gp1ba^−/−;Tg (Gp1ba-Tg) and increased in RhoA^−/− MKs. Scale bar: 10 μm. (d) Quantification reveals decreased Cdc42 activity in Gp1ba^−/−;Tg (light grey) and increased activity in RhoA^−/− (dark grey) polarized MKs compared to the wt (black) (n=4 biological replicates). Bar graphs represent mean±s.d. Two-way ANOVA with Bonferroni correction for multiple comparisons; *P<0.05; ***P<0.001.