Fig. 7: Microglial Tgfb1 deficiency leads to similar microglia-induced demyelination in the DC.

a, Cx3cr1^CreER:Tgfb1^fl/fl mice were employed to delete TGFβ1 production in microglia. b, In 12-month-old Cx3cr1^CreER:Tgfb1^fl/fl mice (D30), a similar demyelinating pathology and microglial activation in the DC were evident, as observed in the Cx3cr1^CreER:Tgfbr2^fl/fl mice. Scale bar, 50 μm. c, t-SNE plots from high-dimensional flow cytometry reveal segregation of the microglia population between Tgfb1^fl/fl and Cx3cr1^CreER:Tgfb1^fl/fl mice (D60). The mapped distribution of F4/80 (green), CD45 (yellow), MHC-II (purple), Galectin-3 (orange) and MSR1 (blue) indicates cells with high marker expression in the t-SNE plots. n = 4 mice per group. d–f, Staining of Galectin-3 (cyan), GPNMB (red) and nuclei (Hoechst, blue) in the VC and DC of Tgfb1^fl/fl and Cx3cr1^CreER:Tgfb1^fl/fl mice (D30) (d), with quantification of Galectin-3 (e) and GPNMB MFI (f). Scale bar, 50 μm; n = 4 and 5 mice for the Tgfb1^fl/fl and Cx3cr1^CreER:Tgfb1^fl/fl groups, respectively. g,h, Comparison of clinical scores (g) and four-paw hanging wire behavioral tests (h) between young (2–3 months; YF) and older female (8–12 months; OF) Cx3cr1^CreER:Tgfb1^fl/fl mice following tamoxifen administration. Tgfb1^fl/fl mice and Cx3cr1^CreER:Tgfbr2^fl/fl mice (OF) were included as negative and positive control groups, respectively. n = 7, 10 and 7 mice for the Cx3cr1^CreER:Tgfbr2^fl/fl (OF), Cx3cr1^CreER:Tgfb1^fl/fl (OF) and Cx3cr1^CreER:Tgfb1^fl/fl (YF) groups, respectively. Data are presented as mean ± s.e.m. *P < 0.05, **P < 0.01 and ***P < 0.001; multiple paired t-tests with FDR correction were used for comparisons between DC and VC in e and f. The cumulative disease score in g was analyzed using the Kruskal–Wallis with Dunnett’s post hoc test, and the mean time in the four-paw hanging wire test in h was analyzed using the one-way ANOVA with Tukey’s post hoc test. Schematic in panel a created with BioRender.com.

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