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. 2020 Jun 10;10:9420. doi: 10.1038/s41598-020-66113-7

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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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Reduced EAAT4 expression on cerebellar Purkinje cells was observed in mice exposed to repetitive blast injury. (a) Shows representative images of secondary only controls (no primary antibodies) for Alexa 488, (b) Alexa Cy3, (c) merged, and (d) zoomed (40×). (e) Shows representative immunofluorescent images of EAAT4 (green), (f) calbindin (red), (g) merged, and (h) zoomed images in lobule IX of the cerebellum in control mice at one month after 3X sham and L-NAME administration. (i) Shows representative images of EAAT4, (j) calbindin, (k) merged, and (l) zoomed images at one month after repetitive mTBI. (m) Shows representative images of EAAT4, (n) calbindin, (o) merged, and (p) zoomed images at one month after repetitive mTBI with L-NAME. A significant decrease in (q) EAAT4 (*p ≤ 0.05), and (r) calbindin (*p ≤ 0.05) immunofluorescence was observed in lobule IX of the cerebellum of mice exposed to both repetitive mTBI plus vehicle (*p ≤ 0.05), and repetitive mTBI plus L-NAME (*p ≤ 0.05). One-way ANOVA post hoc Newman-Keuls. Values represent means ± SEM. Arrowheads highlight EAAT4+/calbindin+ Purkinje cell bodies. Scale bars = 30 µm, 20 µm (zoomed).