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. 2023 Aug 2;620(7973):374–380. doi: 10.1038/s41586-023-06373-1

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© The Author(s) 2023

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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Extended Data Fig. 10 — a, mRNA expression levels of interferon-stimulated genes and activation markers in microglia from Tmem119-creER^T2-cGas^WT/R241E mice 4-OHT-treated or not (n = 3). b, Representative images and quantification of IBA1⁺ microglia B2M intensity from brain sections of Tmem119-creER^T2-cGas^WT/WT and Tmem119-creER^T2-cGas^WT/R241E mice (averaged per mouse, n = 5, from 10–15 cells). Arrows indicate IBA⁺ microglia. Scale bars, 50 μm. c, UMAP plots of sorted hippocampi nuclei’s unsupervised clustering (38 clusters) with identified oligodendrocytes-, astrocyte-, microglia- and neuron-specific gene expression scores. d, Volcano plots of genes differentially expressed between Tmem119-creER^T2-cGAS^WT/WT and Tmem119-creER^T2-cGAS^WT/R241E mice (n = 4) in oligodendrocytes, astrocyte, and microglia MG (FDR ≤ 0.05, Log2FC ≥ 0.3, genes listed in Supplementary Table 6). e, Morris water maze memory test of Tmem119-creER^T2-cGas^WT/WT (n = 6) and Tmem119-creER^T2-cGas^WT/R241E mice (data in Fig. 4h), additionally H-151-treated (n = 5) or not (n = 11), P = 1 × 10⁻⁵. f, Schematic illustrating neuronal cell co-culture experiments. g, Representative images of neurons (MAP2, red) cultured with microglia from Rosa26-creER^T2-cGas^WT/R241E mice (IBA1, green), 4-OHT-treated or not, and aTNF (represents n = 3 experiments). Scale bars, 100 μm. h, Relative survival of MAP2⁺ neurons cultured with conditioned medium from Rosa26-creER^T2-cGas^WT/R241E-isolated macrophages, 4-OHT-treated (n = 6 FOV) or not (n = 4 FOV), and with aTNF (n = 5 FOV), aIFNb (n = 6 FOV, from n = 3 mice) addition, P = 3 × 10⁻⁹. i, Relative survival of MAP2⁺ neurons treated with increasing doses of recombinant TNF (untreated n = 5, 50 ng/ml n = 5, 100 ng/ml n = 4 FOV), from 3 slides per condition, P = 1 × 10⁻⁵. Data are mean ± s.e.m. P values were obtained by one-sided (a) or two-sided (b) unpaired Student’s t-test, one-way ANOVA followed by Tukey’s multiple comparisons tests (h,i) or two-way ANOVA (e).

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