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. 2017 Aug 22;7:9089. doi: 10.1038/s41598-017-09664-6

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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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Thiram inhibits angiogenesis-related mRNA and protein expression. (a–c) The mRNA expression levels of HIF-1a, VEGFA, and VEGFR1 were determined by qRT-PCR. mRNA expression levels from the control group were normalized to 1. GAPDH was used as the loading control. The results are representative of at least three independent experiments, which were run in triplicate, and are expressed as the mean ± SD. Student’s t-test, ^** p < 0.01, ^*** p < 0.001, n = 3. (d–f) The protein expression levels of HIF-1a, VEGFA, and VEGFR1 were determined by Western blotting. (g–i) The results of the western blots were quantified and analyzed using Image-Pro® Plus 6.0. Student’s t-test, ^* p < 0.05, ^** p < 0.01, n = 3; Error bars indicate SEM. ns, not significant. (j,k) The serum concentrations of VEGFA, VEGFR2, and IL-8 were measured using ELISA kits. Student’s t-test, n ≥ 3; Error bars indicate SEM.