Extended Data Fig. 3: Astrocyte epigenetic memory and Ep300 signaling in EAE.
From: Disease-associated astrocyte epigenetic memory promotes CNS pathology
(a) Generation of astrocyte epigenetic memory signature filtered based on adjusted p-value and fold change. Up-signature (red) or down-signature (blue) in astrocytes stimulated twice (2X) than once (1X) are shown. (b) Astrocyte epigenetic memory signature score applied to naive and EAE scRNA-seq astrocyte dataset (Priming, Peak, and Remission)10. (c) Experimental design for (d,e). (d) EAE score for (e) (n = 8 per group). Data shown as mean ± s.e.m. Naïve and EAE induced C57BL/6 mice received ICV administration of IL-1β/TNF (EAE peak, Day 22), and 18-24 h later sorted brain astrocytes were analyzed. (e) qPCR of IL-1β/TNF response of astrocytes (Naïve; n = 12; EAE; n = 8 per group). Representative data of two independent experiments. Unpaired two-sided t-test. (f) Immunostaining (left) and quantification (right) of H3K27ac+ and p300+ astrocytes from sgScrmbl- and sgEp300- transduced mice at 23 days after EAE induction (n = 6 spinal cord sections; n = 3 mice per group). Unpaired two-sided t-test. (g) FluoroMyelin dye staining and percentage of myelin loss in spinal cord from sgScrmbl- and sgEp300-treated mice (n = 9 spinal cord sections; n = 3 mice per group). Lesions indicated by arrowheads. Unpaired two-sided t-test. (h) NF-kB signaling network comparing sgEp300-transduced versus sgScrmbl-transduced astrocytes. (i) Quantification of CNS-resident cells from sgScrmbl- and sgEp300-transduced mice (n = 9 per group). Unpaired two-sided t-test. (j,k) Analysis of CNS T cells (up) and splenic T cells (bottom) from sgScrmbl- or sgEp300-transduced mice (n = 5 per group). Unpaired two-sided t-test. (l) Genome browser snapshots showing the Relb locus. Only regions showing a significant decrease (p-value < 0.05) in accessibility in sgScrmbl-transduced versus sgEp300-transduced astrocytes are highlighted by yellow boxes. (m) Chip-qPCR analysis of p300 recruitment to promoters in primary astrocytes 30 min after IL-1β/TNF stimulation on day 7 (n = 3 per group). Unpaired two-sided t-test. Data shown as mean ± s.e.m.
