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. 2018 Feb 14;8:2996. doi: 10.1038/s41598-018-20448-4

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© The Author(s) 2018

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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VPS35 KD does not affect SypHy release or retrieval. (a) Representative sypHy fluorescence images of neurites during baseline, stimulation, the recovery period and the exposure to NH₄Cl. Scale bar = 10 µm. (b) Time course of sypHy fluorescence during the imaging protocol, plotted as ΔF/Fmax. The grey box indicates the electrical stimulation (100 AP, 40 Hz, 30 mV) and the black box the exposure to 10 seconds of NH₄Cl (n = 23 ± 8 fields of view, N = 4 animals). (c) Maximum response amplitude during the electrical stimulation plotted as ΔF/Fmax. (d) Percentage of responsive synapses during the stimulation. (e) Maximum response to the exposure to NH₄Cl. Detailed information (average, SEM, n and statistics) is shown in Supplementary Table S1.