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. 2017 Dec 4;7:16875. doi: 10.1038/s41598-017-17061-2

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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 signaling is altered in GEC isolated from Alport mice. VEGF signaling changes in GEC were studied using tdTomato-labeled GEC (tdT GEC) isolated by FACS from Alport-Tek^tdT glomeruli (A, 10x). tdT signal (red) is strongly present in all cells as shown in (B) (10x, two passages in culture) (n = 3). Graphs representing immunoblot data of pAKT/AKT (C, 60 kDa/60 kDa), pERK/ERK (D, 42 kDa/42 kDa), VEGFR1 expression (E, 151 kDa) in glomeruli of tdT GEC derived from WT mice and Alport-Tek^tdT mice at different ages. Imbalanced VEGF signaling is evident during disease progression in GEC, identified by a strong alteration in pAKT/AKT and pERK/ERK signaling downstream of VEGFR2. (F) Immunoblots of all the experimental groups used to generate the graphs presented in Figures C–E. Immunoblots were quantified by densitometry (VEGFR1 measurements were normalized against corresponding housekeeping gene, β-actin, 42 kDa). These data were obtained using GEC derived from n = 4 WT at 5 months of age, n = 4 AS mice at 2 months of age, n = 6 AS mice at 3 month of age, n = 4 AS mice at 6 months of age. One-way ANOVA with Tukey’s post hoc test was used to analyze the data and scatter plot values are presented as mean ± SD (*p < 0.05).