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. 2017 Aug 23;7:9216. doi: 10.1038/s41598-017-10005-w

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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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Pathways integral to vascular homeostasis appear unperturbed in mutant Gars muscle. (A) Representative Nrp1 (white) and VEGFR2 (red) staining in one month old wholemount wild-type and Gars ^C201R/+ TVA muscles. Nrp1 and VEGFR2 colocalise with the endothelial marker IB4 (green) in both genotypes, and show no obvious differences. A similar pattern was seen at three months and in the lumbrical muscles at both time points (data not shown). Both panels are single plane images. Scale bars = 200 μm. (B,C) qPCR analysis of a series of genes integral to the maintenance of blood vessels indicates very little perturbation in gene expression in one month Gars ^C201R/+ tibialis anterior muscles. This was confirmed using either Gapdh (top graphs) or endothelium-specific Pecam1 (bottom graphs) as the reference gene. All individual gene data sets were separately analysed with unpaired, two-tailed t-tests. n = 6 wild-type and 4 Gars ^C201R/+.