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. 2020 Jul 21;10:12057. doi: 10.1038/s41598-020-68970-8

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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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ImageJ composites illustrative of the count of presynaptic boutons expressing SV2B (green), CLT (red), or both immunoreactivities in control (A) and tbl (B) hippocampal cultures. (C–E), graphs showing the number of single SV2B and double (SV2B-CLT) labelled boutons (C), the percentage of labelled boutons (D), and their density of counted boutons per dendrite µm (E). The number of presynaptic boutons is higher in control relative to tbl cultures (C, T: *p = 0.014; SV2B-CLT: **p = 0.005; SV2: *p = 0.04). Correlative to these counts, the percentage of double labelled boutons is lower in tbl cultures relative to control ones (D, SV2B-CLT: **p = 0.007). The density of the number of all categories of presynaptic boutons counted is ever lower in tbl cultures than in control ones (E, T: **p = 0.005; SV2B-CLT: **p = 0.003; SV2: **p = 0.009), while no significant differences were found on the mean of the dendrite’s length measured (F; p = 0.544). Bars = 5 µm. T, total number of counted boutons.