Fig. 3

Proliferative microglia cluster emerged in the ischemic mice brain. A t-SNE representation of MG4 (left); violin plots showing the top up-regulated genes in MG4 (right). B Feature plots displaying cell cycle score determined by expression of highly conserved cell cycle genes. C, D Representative confocal images of MKI67 and IBA-1 double immunostaining in the ischemic penumbra and quantification of the MKI67⁺/IBA‐1⁺ cells at different timepoints after tMCAO. Scale bar: 40 μm (overviews). Triangles indicate resident MKI67⁺ microglia. one-way ANOVA with Bonferroni multiple comparisons test. n = 6–8. The data are shown as means ± SD. *p < 0.05, ***p < 0.001, n.s. no significance. E, F Representative confocal images of CH25H/OASL, IBA-1 and EdU treble immunostaining in the ischemic penumbra 3d after tMCAO. (tMCAO transient middle cerebral artery occlusion, t-SNE t-distributed stochastic neighbor embedding, MG microglia, IBA-1 ionized calcium binding adapter molecule 1, ANOVA analysis of variance.)