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. 2017 Nov 7;7:14749. doi: 10.1038/s41598-017-15246-3

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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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VEGF activated GlyT1 and glycine directly bound to VDAC-1 on the mitochondria outer membrane. (a) Immunoblots of SHMT1 and SHMT2 in ECs treated with VEGF (n = 3). (b) The effects of sarcosine, an inhibitor of glycine transporter on the proliferation of ECs treated with VEGF (n = 12). (c) Immunoblots of GlyT1 and GlyT2 in ECs treated with VEGF (n = 3). (d) GlyT1 protein expression by Western blots in HUVECs electroporated with GlyT1 siRNA (si-GlyT1) or control siRNA (si-nc) treated with 20 ng/ml VEGF. (e) LC-MS was used to detect relative intracellular glycine in ECs with si-GlyT1 in the absence and presence of VEGF (n = 3). (f) VDAC1 was reconstituted in the plasma membrane of Hek-293A. The effects of glycine on single channel currents of VDAC-1 were recorded in +40 mV voltages using single channel patch-clamping (n = 7). (g) Statistical analysis of open probability (NPo) in the presence of Gd3 +(a VDAC1 inhibitor) and glycine (n = 7). All values represented mean ± SEM of n independent experiments. **P < 0.01, 1-way ANOVA.