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. 2017 Aug 7;7:7393. doi: 10.1038/s41598-017-07952-9

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© The Author(s) 2017

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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HIMF promotes VEGF expression and angiogenesis in vivo. (A) Matrigel plugs containing myoblast conditioned medium (CM) of PBS (vehicle), HIMF (10 nM), or anti-IL-18 antibody combined with HIMF were subcutaneously injected into the flanks of nude mice. After 7 days, the plugs were photographed. (B) Hemoglobin levels from the plugs were quantified and normalized to control group. (C) Specimens from the plugs were immunostained with antibodies against CD31 and VEGF. Scale bar: 100 μm. (D) Data from a CAM assay using 5-day-old fertilized chick embryos (6 eggs/group): PBS (vehicle), CM with HIMF (10 nM) treatment, or CM with anti-IL-18 antibody followed by HIMF treatment were resuspended in Matrigel and placed onto the CAMs, which were allowed to develop for another 3 days. The CAMs were then examined by microscopy and photographed. Scale bar: 2 mm. (E) CAM vessels were calculated and quantified. Results are expressed as the means ± SEM of four independent experiments. *p < 0.05 as compared with the control group. #p < 0.05 as compared with HIMF-treated group.