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. 2024 Jan 10;15:449. doi: 10.1038/s41467-024-44704-6

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Schematic of bilateral virus infection. Representative images of neuronal dendrites (b) and quantification of spine numbers per 10 μm (c) (n = 6 mice per group; F_1,30 = 31.40, p < 0.001). Scale bars, 10 μm. d Experimental scheme for virus injection. Representative images of neuronal dendrites (e) and quantification of spine numbers (f) from the indicated groups (n = 6 mice per group; f F_1,20 = 7.743, p = 0.0115). Scale bars, 10 μm. g Representative images (left) and quantitative analyses (right) of immunostaining for CD68 (red), Iba1 (green), and DAPI (blue) in the CeA from the indicated groups (n = 6 mice per group; Iba1, F_1,20 = 91.50, p < 0.001; CD68, F_1,20 = 30.28, p < 0.001). Scale bars, 20 μm. h Representative images and three-dimensional surface rendering (left) of Iba1+ microglia (green) containing GAD65/67+ puncta (red) and DAPI (blue) in the CeA from the indicated groups. Quantification (right) of GAD65/67⁺ puncta in microglia in mice (n = 53 cells from six mice per group; F_1,208 = 4.789, p = 0.0298). Scale bars, 10 μm (overview) and 5 μm (inset and rendering). i Representative images (left) and quantitative analyses (right) of immunostaining for CD68 (red), Iba1 (green), and GAD65/67⁺ puncta (purple) in the CeA from the indicated groups (n = 24 cells from six mice per group; F_1,92 = 26.55, p < 0.001). Scale bars, 5 μm. j Representative images (left) and quantification (right) of Iba1⁺ microglia (green) containing GAD65/67⁺ puncta (red) in the CeA from the indicated groups (n = 30 cells from six mice per group; t₅₈ = 6.739, p < 0.001). Scale bars, 10 μm (overview) and 5 μm (inset and rendering). Significance was assessed by two-way repeated-measures ANOVA with post hoc comparison between groups in (c, f, g, h, i), two-tailed unpaired Student’s t-test in (j). All data are presented as mean ± SEM. *p < 0.05, ***p < 0.001; n.s., not significant. See also Supplementary Data 1. Source data are provided as a Source Data file.