Extended Data Fig. 2: Regulation of HB-EGF by astrocytes during autoimmune neuroinflammation.

a, EAE development in wild-type mice with the timepoints analyzed (naïve, peak, recovery, LSW) indicated. n = 20. b, tSNE plots of CNS cells (upper, downsampled to 30000 cells) and cell type abundance (% of singlets; c) analyzed by high-dimensional flow cytometry. OLCs, oligodendrocyte lineage cells; OPCs, oligodendrocyte precursor cells; NLCs, neuronal lineage cells; Endo, endothelial cells; DCs, dendritic cells; Macro, macrophages. Naïve n = 5, Peak n = 6, Recovery n = 4, LSW n = 8. d, median fluorescence intensity (MFI) of CXCL-12, TNF-α, and Ki67 in astrocytes during peak and late-stage worsening (LSW) quantified by intracellular flow cytometry. n = 3 per group. e, principal component analysis (PCA), heatmap (f), and volcano plot (g) of differential gene expression in ACSA2⁺ astrocytes during peak and LSW analyzed by RNA-Seq. n = 3 per group. h, KEGG Pathway enrichment of LSW astrocytes. i, relative expression (% of parent, left) and MFI (right) of HB-EGF production by cortical and spinal cord astrocytes in naïve (n = 3) and EAE mice (n = 5). j, UMAP plot of ACSA2⁺ cells (downsampled to 5000 cells) in naïve and EAE mice analyzed by high dimensional flow cytometry. Data shown as mean ± SD if not indicated otherwise. Data shown as mean ± SEM in (a). Two-way ANOVA with Tukey’s multiple comparisons test in (c), unpaired t-test with Holm-Sidak correction in (d), Two-way ANOVA with Sidak’s multiple comparisons test in (h).

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