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. 2020 Jan 28;10:1349. doi: 10.1038/s41598-020-58452-2

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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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Apelin protected vascular reduction and suppressed hypoxia and apoptosis after cutaneous I/R. (A) The amount of CD31⁺ ECs and NG2⁺ pericytes in the cutaneous I/R area at 4 days after reperfusion. (B) The amount of pimonidazole⁺ hypoxic area in cutaneous I/R site at 1 day after reperfusion. Quantification of the pimonidazole⁺ areas in 6 random microscopic fields from the center of I/R area in n = 3 mice per groups was performed using ImageJ software. Positive area in control mice was assigned a value of 1. Scale bar = 20 μm. (C) The number of apoptotic cells in the I/R site at 1 day after reperfusion was determined by counting both TUNEL- and DAPI-positive cells. Values were determined in 6 random microscopic fields in n = 3 mice per group. All values represent mean ± SEM. **P < 0.01, *P < 0.05.