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. 2020 Jan 22;10:968. doi: 10.1038/s41598-020-57929-4

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© The Author(s) 2020

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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Effects of SYN1 R420G mutation on hippocampal neurons. (A) Western blot in HEK293T cells untransfected or transfected with either wild-type of R420G SYN1-V5 mutant confirm expression of both constructs. Original western blot images are provided as Supplementary Material (B) Transfection efficiency on hippocampal neurons was calculated in three independent experiments by counting GFP+ cells. Black solid column shows wild-type SYN1-V5 transfected neurons and solid grey column shows R420G SYN1-V5 mutant transfected neurons C: Microscopy images of hippocampal neurons transfected with either wild-type (upper panel) or mutant (lower panel) SYN1-V5 (Scale bar: 25μM). (D) Neurons transfected with wild-type SYN1-V5 express higher levels of SYN1 (p = 0.0004) and (E) have longer axons (p = 0.0008) than their mutant counterparts. The length of axons and normalized fluorescence were measured using the ImageJ software. The graphs shows the mean normalize fluoresce (upper graph) and the axon length in μM (lower graph) of three independent transfections. Black columns show wild-type SYN1-V5 transfected neurons and solid grey columns show R420G SYN1-V5 mutant transfected neurons. Values represent the means ± SEM. ***p < 0.001; ns = non-significant.