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. 2018 Jan 17;8:879. doi: 10.1038/s41598-018-19197-1

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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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Effects of ILO and ILO-PC on rapid and sustained enhancement of adherens junctions. (A) EC monolayers were incubated with 0.5 µM ILO or ILO-PC for 15 or 90 min. Increased areas of adherens junctions in response to agonist treatment were monitored by immunofluorescence staining of transmembrane adherens junction protein, VE-cadherin. Bar = 10 µm. Insets with higher magnification images show details of adherens junction structure. (B) Membrane accumulation of adherens junction proteins VE-cadherin and p120-catenin after 90-min stimulation with 0.5 µM ILO or ILO-PC detected by subcellular fractionation assay. (C) Membrane accumulation of VE-cadherin and p120-catenin caused by ILO or ILO-PC was analyzed in the presence or absence of Rac1 and Rap1 pharmacological inhibitors (30-min pre-treatment with 100 µM NSC23766 and 10 µM GGTI298, respectively). Probing of total lysates for VE-cadherin and p120-catenin was used as a normalization control. Bar graphs depict quantitative densitometry analysis of western blot data; n = 4; *p < 0.05. Data are expressed as mean ± SD. Biological replicates indicated by (n). Statistical significance by one-way analysis of variance (ANOVA) and Tukey’s post hoc multiple-comparison test.