Extended Data Fig. 6: Astrocyte epigenetic memory in NOD EAE.

(a) Experimental design for (c). (b) NOD EAE score for (c) (n = 3 per group). (c) Naïve and EAE induced NOD mice received ICV administration of IL-1β/TNF (EAE progressive, Day 124). After 18-24 h, sorted brain astrocytes were analyzed by qPCR (n = 3 per group). Unpaired two-sided t-test. (d) Immunostaining (left) and quantification (right) of H3K27ac⁺ and p300⁺ astrocytes in mice with/without NOD EAE (n = 9 spinal cord sections; n = 3 mice per group). Astrocyte H3K27ac levels were calculated as the mean signal intensity (arbitrary units) per GFAP+ cells using automated unbiased quantification. Unpaired two-sided t-test. (e) Immunostaining (left) and quantification (right) of ACLY⁺, p300⁺, and ACLY⁺p300⁺ astrocytes in mice with/without NOD EAE (n = 9 spinal cord sections; n = 3 mice per group). Unpaired two-sided t-test. (f) NOD EAE curves (sgScrmbl; n = 7; sgEp300; n = 8; sgAcly; n = 7). Lentivirus were injected at day 40. Representative data of two independent experiments. Regression slope two-sided t-test compared with sgScrmbl. (g) Volcano plot of differential gene expression determined by RNA-seq in astrocytes isolated from sgScrmbl-, sgEp300-, and sgAcly-transduced mice 64 days after NOD EAE induction (n = 3 sgScrmbl, n = 3 sgEp300, n = 2 sgAcly). (h) GSEA analysis comparing sgScrmbl-, sgEp300-, and sgAcly-transduced astrocytes. (i) Staining with FluoroMyelin dye and percentage of myelin loss from sgScrmbl-, sgEp300-, and sgAcly- transduced mice spinal cord (n = 6 spinal cord sections (sgEp300); n = 9 spinal cord sections (sgScrmbl, sgAcly); n = 3 mice per group. Lesions indicated by arrowheads. Unpaired two-sided t-test. Data shown as mean ± s.e.m.

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