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. 2017 Jun 22;7:4092. doi: 10.1038/s41598-017-04460-8

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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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Neutralization of BDNF reduces DRG axonal growth in direct co-culture with HUVECs. DRG explants were cultured on (A) HUVECs and (B) HUVEC cultures supplemented with 125 ng/mL of BDNF neutralizing antibody. DRG-HUVEC co-cultures without neutralizing antibody displayed asymmetrical growth patterns, with long axons growing towards clusters of HUVECs (arrows). Supplementation of cultures with neutralizing antibodies restored symmetrical growth patterns. Scale = 200 µm. (C) Quantification of DRG axonal outgrowth from explants cultured with high or low density of HUVECs and with or without BDNF neutralizing antibodies. Neutralization of BDNF significantly reduced axonal outgrowth, while low density seeding conditions supported more axonal outgrowth independent of the addition of the neutralizing antibody. Data are presented as mean ± standard error. Brackets (p < 0.001) indicate significance between HUVEC densities and ***(p < 0.001) indicate significance between cultures with and without neutralization antibodies as determined by two-tailed Student’s t-tests (DRG sample size for each group shown in bars from 4 independent replicates).