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. 2017 May 19;7:2149. doi: 10.1038/s41598-017-02202-4

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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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Survival is altered in low-density cultures of EndophilinA triple knockout neurons. (A,B) Brain sections of wild-type and EndophilinA triple knockout mice immunostained with Neurotrace (green) and DAPI (red) showed no obvious defects in overall structure of the hippocampus or the number of Neurotrace/DAPI-positive cells (quantified in C,D). (E) High-density neuronal cultures of wild-type and EndophilinA triple knockout neurons immunostained for acetyl tubulin. Both wild-type and EndophilinA TKO neurons formed networks and survived up to 14 days in culture. (F) Survival assay of wild-type and Endophilin A1-/-; A2-/-; A3-/- low-density cultures: EndophilinA TKO neurons showed higher rates of cell death compared to wild-type neurons after 48 hours in culture in the absence of BDNF. This increase in cell death in Endophilin A1-/-; A2-/-; A3-/- neurons could not be rescued by administration of 100 ng/ml exogenous BDNF compared to control during a 120-hour time course; n = 4 independent experiments, significance determined by unpaired two-tailed Student’s t-test, comparing each condition to numbers of initially plated neurons. Error = SEM, *p < 0.05, **p < 0.01.