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. 2018 Jun 22;8:9575. doi: 10.1038/s41598-018-27810-6

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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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Estrogenic condition regulates HUVEC viability, motility and tube formation via CCL2-CCR2 axis in vitro. (a) The proliferation of HUVECs that were supplied with or without different concentrations of rhCCL2 or DMSO/E2-treated CM in the present or absence of RS102895 (20 μM) at the indicated time was measured by WST-8 assay; (b) HUVECs were supplied with or without 50 ng/ml rhCCL2 or DMSO/E2-treated CM from MCF-7 and T47D cells in the pretreatment or absence of RS102895 (20 μM) and their migratory ability at 24 h post incubation was determined by migration assay; (c) HUVECs were treated with or without various concentrations of rhCCL2 in the pretreatment or absence of RS102895 (20 μM) and their ability to form capillary-like tubes were measured using tube formation assay. (d) HUVECs were incubated in the presence of DMSO-treated or E2-treated CM from MCF-7 and T47D cells with the pretreatment of RS102895 or not. Then HUVECs were captured 10 hours after CM was added. Images are representative and data are shown as the mean ± SEM from three separate experiments. *P < 0.05, **P < 0.01, ***P < 0.001 vs. control or DMSO CM; ns, not significant.