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. 2018 May 15;8:7595. doi: 10.1038/s41598-018-25874-y

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© The Author(s) 2018

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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NGR1 prevents hCASMC migration through reorganization of actin cytoskeletal dynamics. (A) Representative images of quiescent hCASMCs that were pretreated with NGR1 for 24 h and stimulated with serum for 24 h. Images were captured at 0, 16, 24 h. Quantification of percentage of area covered by migrated cells over time was calculated using ImageJ (right panel). N = 3. Scale bar, 50 μm. (B) Representative images from transwell migration assay with different conditions (left panel). Quantification for the number of migrated cells are shown in the bar graph on the right. N = 3. (C) Representative images of F-actin (Alexa-488-tagged phalloidin), G-actin (Alexa-594-tagged DnaSe1) and DAPI stained hCASMCs that were unstimulated (Con) or treated with FBS or FBS + NGR1 (10 µM). Lamellipodia are indicated by the white arrows. Scale bar, 2 μm. (D) Quantitative results show the ratio of F-actin and G-actin on the basal plane of hCASMC images. Fifty cells in each condition were quantified. All experiment was repeated at least 3 times. Data shown are means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 vs. serum control group.