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. 2018 Oct 26;8:15885. doi: 10.1038/s41598-018-34357-z

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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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HUVEC release cardioprotective factors. (A) Experimental protocol for the co-culture of HUVEC with adult rat cardiomyocytes using a trans-well system, followed by hypoxia and reoxygenation (H/R). Separate wells of cardiomyocytes were maintained in normoxic medium. Insulin pre-treatment was used as a positive control for cardioprotection. (B,C) Representative images of cardiomyocytes stained with propidium iodide (PI) at the end of the H/R protocol. Fluorescent images (PI staining in red) were overlaid with phase contrast images of cells. (D,E) Percentage of dead cells after normoxic culture, after H/R, after HUVEC co-culture (D, N = 5), or after addition of exosome-free conditioned medium (“exo-free CM”) (E, N = 3). *P < 0.05, ***P < 0.001.