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. 2024 Apr 3;15:2866. doi: 10.1038/s41467-024-46625-w

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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. 7 — a, b UMAPs illustrating subclusters of microglia (a) and cell distributions (b) among those subclusters at all time points (intact, 3 dpi, and 5 dpi) and conditions (CTRL and INH). Data shown in (b) are downsampled to an equal number of cells between time points and conditions. Data shown in panels (a) and (b) are derived from n = 12 intact animals over 3 independent experiments (5 scRNAseq libraries), n = 3 3dpi CTRL animals over 1 experiment (2 scRNAseq libraries), n = 3 3dpi INH animals over 1 experiment (2 scRNAseq libraries), n = 9 5dpi CTRL animals over 3 independent experiments (3 scRNAseq libraries) and n = 6 5dpi INH animals over 2 independent experiments (3 scRNAseq libraries). c Experimental paradigm to assess microglial reactivity. Gray boxes on the mouse brain schemes highlight the analyzed areas. Red lines indicate the injury cores. d–g Plots depicting the distribution of microglial circularity (d), soma volume (e), branch volume (f), and number of nodes per major branch (g) of n_CTRL = 6 and n_INH = 5 animals. Source data are provided as Source Data file. Data are fitted with multiple peak functions. The fit parameters are depicted in Supplementary Data 8 for each fit. h, i Representative images of P2Y12 staining in CTRL (h) and INH-treated (i) mice. Dashed white lines indicate the injury core. All images are full z-projections of confocal z-stacks. j Dot plot depicting the mean gray value of P2Y12⁺ signal in the injury vicinity of CTRL and INH-treated mice at 5 dpi. Data are shown as mean ± standard error of the mean. Each data point represents one animal. Source data are provided as Source Data file. Statistics have been derived from n_CTRL = 6 and n_INH = 7 animals. p-values were determined with unpaired t-test (two-tailed). k Violin plots depicting expression levels of P2ry12 in microglia (scRNA-seq) in intact, 5 dpi CTRL and 5 dpi INH condition. Data are derived derived from n = 12 intact animals over 3 independent experiments (5 scRNAseq libraries), n = 9 5dpi CTRL animals over 3 independent experiments (3 scRNAseq libraries) and n = 6 5dpi INH animals over 2 independent experiments (3 scRNAseq libraries). Scale bars: h, i: 50 μm. UMAP uniform manifold approximation and projection, dpi days post-injury, CTRL stab wound-injured control animals, INH stab wound-injured inhibitor-treated animals, INT intact.