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. 2021 Sep 16;19:95. doi: 10.1186/s12964-021-00773-4

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© The Author(s) 2021

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 6 — DN R-Ras decreases phosphorylation of ROCK1 and alters migration and cell morphology. a Effect of DN R-Ras expression on the actin cytoskeleton in ST88-14 cells as assessed by phalloidin staining. The enlarged image demonstrates actin-rimmed vesicular structures (white arrows) that accumulate in ST88-14 cells following induction of DN R-Ras expression. b, c Total ROCK1 (b; red) or phospho-ROCK1 immunoreactivity (c; red) in ST88-14 MPNST cells in the presence (+) or absence (−) of DN R-Ras expression and treatment with vehicle (−) or the ROCK inhibitor Y27632 (+). Cells were counterstained with phalloidin counterstain (green) to demonstrate cell morphology as highlighted by the actin cytoskeleton and bisbenzimide (blue) to label nuclei. Arrows denote notable changes in the actin cytoskeleton, including a loss of filamentous structures and the accumulation of actin-rich puncta (white arrows) following induction of DN R-Ras expression or Y27632 treatment. A non-immune isotype matched primary antibody was used as a negative control. Scale bars = 25 μm. d Semi-quantitative analysis of normalized total and phosphorylated ROCK1 levels relative to uninduced or untreated controls. **, p-value < 0.0001