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. 2019 Dec 2;9:18143. doi: 10.1038/s41598-019-54211-0

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© The Author(s) 2019

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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Influence of GAG derivatives on VEGF₁₆₅-mediated phosphorylation of VEGFR-2 (a,b) and direct interaction of solute GAG derivatives with immobilized VEGFR-2 (c). (a) HUVEC cells were stimulated with GAG for 10 min. Cells were lysed afterwards and lysates were applied to a specific phospho- and a total VEGFR-2 sandwich ELISA. The amount of phospho-VEGFR-2 relative to total VEGFR-2 is shown. Values represent the mean ± SD of n = 2. One-way ANOVA: ^#p < 0.05 vs. Ctrl; *p < 0.05 vs. VEGF₁₆₅. (b) HUVEC cells were stimulated with VEGF₁₆₅ alone or pre-formed VEGF/GAG complexes for 10 min. Cell lysates were applied to a specific phospho- and a total VEGFR-2 sandwich ELISA. The amount of phospho-VEGFR-2 relative to total VEGFR-2 is shown. SU1498, a specific VEGFR-2 inhibitor was used as a reference for reduced phosphorylation. Values represent the mean ± SD of n = 3. One-way ANOVA: ^#p < 0.05 vs Ctrl; **p < 0.01 vs. VEGF₁₆₅ alone. (c) One representative sensorgram out of three independent measurements is shown [adapted and modified from³³].