Fig. 4: Spinal cord snRNA-seq reveals a TSM subset.
From: TGFβ signaling mediates microglial resilience to spatiotemporally restricted myelin degeneration
a, A UMAP plot of spinal cord microglia from 12-month-old female Tgfbr2fl/fl (WT) mice and Cx3cr1CreER:Tgfbr2fl/fl (KO) mice at D10, D20 or D30 post-tamoxifen administration (downsampled to 500 cells per group per time point for presentation). b, The kinetic fold change (compared to WT condition) of each microglial subcluster in a over time. c, The proportion of each microglial subcluster in the total microglial population in WT and KO_D30 conditions. Subclusters 0, 1, 2 and 3 in a and b are denoted as MyTE, TSM, TRM or homeostatic microglia (HM), respectively. d, A bubble plot depicting the expression of marker genes across different subclusters. Bubble size represents the percentage of cells expressing each gene within a subcluster and color indicates the average normalized expression level. Marker genes were identified using Seurat’s FindAllMarkers function (two-sided Wilcoxon rank-sum test with Benjamini–Hochberg correction for multiple comparisons). e, GO enrichment analysis based on the upregulated DEGs in TSM compared to other subclusters; log2 fold change >1, adjusted P value <0.001. f–h, Immunostaining validation of TSM signature markers GPNMB and Galectin-3 (Lgals3) in the spinal cords of 12-month-old WT and KO mice (scale bar, 100 or 20 μm), with quantification of GPNMB and Galectin-3 MFI (h) and insets in g showing costaining of F4/80–GPNMB and IBA1–Galectin-3. n = 6, 3, 3, 5 and 5 mice for each group. Data are presented as mean ± s.e.m. *P < 0.05 and **P < 0.01; multiple paired t-tests with FDR correction.
