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. 2020 Jul 2;10:10913. doi: 10.1038/s41598-020-67988-2

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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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Protease defective TeNT does not block DCV fusion. (A) Representative images of DIV 14 neurites infected (DIV 10) with NPY-pHluorin and control, TeNT or TeNT-E234Q, stained for VAMP2 and the HA-tag. Only TeNT-E234Q was N-terminally tagged with HA. Merge indicates overlay of VAMP2 (green) and HA-tag (magenta) only. (B) Histograms of DCV fusion events in control (black), TeNT (red) or TeNT-E234Q (purple) infected hippocampal neurons (infected with NPY-pHluorin at DIV 9 and control, TeNT or TeNT-E234Q at DIV 10, imaged at DIV 14). Blue bars indicate 16 trains of 50 AP at 50 Hz interspaced by 0.5 s. (C) Cumulative plot of DCV fusion events of conditions as in B. Shaded area represents SEM. (D) Average DCV fusion events per cell for control (n = 19, N = 2), TeNT (n = 11, N = 2) and TeNT-E234Q (n = 18, N = 2) infected neurons. Kruskal–Wallis with Dunn's correction: ***p < 0.10* 10⁻³, non-significant (ns) p > 0.05. Bars represent mean + SEM. Detailed statistics are shown in Supplementary Table S1.