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. 2017 Nov 13;7:15386. doi: 10.1038/s41598-017-13071-2

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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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Increased Eph-B4 and Ephrin-B2 expression during adaptive venous remodeling. (A) Western blot and adjacent bar graph of densitometry showing human Eph-B4 expression in AVF venous limb compared to normal vein. *P = 0.0016; t-test. n = 3–4. (B) Line graphs show expression of Eph-B4 (blue) and Ephrin-B2 (red) in the AVF venous limb compared to sham IVC; P < 0.0001 (ANOVA). *P < 0.05 (P = 0.0123, Eph-B4; P = 0.0041, Ephrin-B2; post hoc); **P < 0.05 (P < 0.0001, Ephrin-B2; post hoc). n = 5–8. (C) Western blots showing Eph-B4 and Ephrin-B2 protein expression in AVF venous limb compared to sham IVC. n = 3–5. (D) Graphs showing densitometry of Eph-B4 (left panel) and Ephrin-B2 (right panel) expression in the AVF venous limb compared to sham IVC; *P < 0.05 (P < 0.0001, Eph-B4 day 7, AVF vs sham; P < 0.0001, Eph-B4 day 21, AVF vs sham; P < 0.0001, Ephrin-B2 day 7, AVF vs sham; post hoc). n = 3–5. (E) Diagram of rat model showing location of infrarenal IVC pericardial patch exposed to an aortocaval AVF (n = 6 per group). (F) Representative Western blot (upper panel) showing Eph-B4 and Ephrin-B2 expression in patch neointima (day 14) of control vein compared to patch neointima of AVF vein. Graphs (lower panel) show quantification of western blot bands; P < 0.0001 (ANOVA). *P < 0.05 (P = 0.0003, Eph-B4; P = 0.0043, Ephrin-B2; post hoc). n = 3. (G) Representative photomicrographs (upper panel) showing Eph-B4 (green) and Ephrin-B2 (red) immunoreactive signal (day 14). White arrowheads indicate colocalization of Eph-B4 and Ephrin-B2. L, vessel lumen. Graph (lower panel) shows quantification of immunoreactive signal; P < 0.0001 (ANOVA). *P < 0.05 (P = 0.0136 Eph-B4; P < 0.0001 Ephrin-B2; post hoc). n = 3. Scale bar 100 µm. Data represent mean ± SEM.