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. 2020 Oct 29;11(10):931. doi: 10.1038/s41419-020-03132-2

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© The Author(s) 2020

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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Fig. 5 — a Phalloidin staining of actin cytoskeleton in HaCaT cells transfected with RNA duplexes as indicated at 24 h post TGF-β1 treatment. Representative images and quantitative data of lamellipodia versus cell surface showed attenuated lamellipodia formation in miR-200b/c-3p or siRAC1-transfected cells (lamellipodia versus cell surface, n = 6). b HaCaT cells were transfected with RNA duplexes as indicated and confluent cells were scratch wounded and treated with TGF-β1 for 6 h. RAC1 protein was shown by immunofluorescence staining (green) and actin cytoskeleton was shown by phalloidin staining (red). Control cells in the leading edge form lamellipodia and RAC1 protein level increased and accumulated in the membrane ruffles facing the migrating direction. RAC1 in miR-200b/c-3p-transfected cells remained in perinuclear area with slight elevation in the leading edge. These cells seldom form lamellipodia. siRAC1 significantly reduced RAC1 indicated by immunofluorescence average integrated optical intensity (IOD) and prevented cells from lamellipodia formation (n = 12). c HaCaT cells treated as in (b) were fixed at 24 hpw. Leader cells transfected with miR-200b/c-3p or siRAC1 became elongated. Stress fibers (indicated by arrowheads) were more obvious in miR-200b/c-3p or siRAC1-transfected cells than control in follower cells. Data presented as mean ± SD. **P < 0.01, ***P < 0.001 versus negative control, one-way ANOVA. Scale bars: 25 μm.