Fig. 3 |. MAFG in astrocytes promotes CNS inflammation.

a, K-means clustering by region of expression data from Ribotag^Gfap astrocytes projected onto cluster 5 (C5) TdTomato^Gfap Nfe2l2 IPA network. Naive, n = 3; peak: n = 3 for fourth and fifth columns, n = 2 for first, second and third columns; remission: n = 3 for second, fourth and fifth columns, n = 2 for first and third columns. First column, cerebellum; second, cortex; third, cranial nerves; fourth, parenchyma; fifth, spinal cord. N, naive; P, peak; R, remission. b, Western blot (top) and quantification of induction (bottom left) of MAFG expression in activated astrocytes. Kruskal–Wallis test, uncorrected Dunn’s test. qPCR analysis (bottom right). n = 6 independent biological samples per condition, unpaired two-tailed t-test. c, d, Immunostaining (c) and quantification (d) of MAFG⁺NRF2⁻ astrocytes in mice with and without EAE (naive). Left, per cent MAFG⁺NRF2⁻ astrocytes. Spinal cord (SC), n = 117 cells from naive mice, n = 260 from mice with EAE; corpus callosum, n = 112 naive, n = 277 EAE; cortex, n = 27 naive, n = 67 EAE; χ² test. Right, number of MAFG⁺NRF2⁻ astrocytes. n = 13 sections for spinal cord naive, n = 9 sections otherwise, from n = 3 mice; unpaired two-tailed t-test. e, ChIP–qPCR analysis of MAFG recruitment to the Hmox1 promoter, n = 5 per group. One-sample t-test on log-normalized data. f, Antioxidant response element (ARE) motif analysis by ChIP–seq (day 23 after induction of EAE). Panels denote ARE motif sequences. n = 3 mice per group. g, EAE progression in mice transduced with sgScrmbl (n = 25) or sgMafg (n = 15). Top: schematic of lentiviral vector. Experiment repeated three times. Two-way repeated measures ANOVA. h, i, RNA-seq analysis (h) and GSEA (i) of gene expression in astrocytes from mice transduced with sgScrmbl or SgMafg. n = 3 mice per group (day 22). Data shown as mean ± s.e.m.