Fig. 3: Neuroectodermal IL-12-sensing attenuates CNS immunopathology.

a, Schematic of Nestin^Cre/+Il12rb2^fl/fl and Il12rb2^fl/fl control mice. b, Il12rb2 expression in FANS-isolated Hoechst⁺Olig2⁺ oligodendrocyte and Hoechst⁺NeuN⁺ neuronal nuclei from Nestin^Cre/+Il12rb2^fl/fl (n = 7, m/f) and Il12rb2^fl/fl control mice (n = 7, m/f) at peak EAE (13 d.p.i.). Data are pooled from two experiments (two-tailed Mann–Whitney test, ***P = 0.0006; ***P = 0.0006, left to right). c,d, EAE course (c) and maximal EAE scores (d) of Nestin^Cre/+Il12rb2^fl/fl (n = 8, m/f), Il12rb2^fl/fl littermate controls (n = 10, m/f), Il12rb2^del/del (n = 7, m/f) and Nestin^Cre/+Il12rb2^+/+ (n = 9, m/f) mice. Data were pooled from two experiments. Mixed-effects model with Bonferroni post hoc test between Il12rb2^fl/fl and Nestin^Cre/+Il12rb2^fl/fl mice (*P = 0.0209; **P = 0.0045; *P = 0.0288; **P = 0.0046; ***P = 0.0005; **P = 0.0092; *P = 0.0221; **P = 0.0014; *P = 0.0131; left to right) in c and two-tailed Mann–Whitney test (P > 0.9999; ****P < 0.0001; **P = 0.0022, left to right) in d. e, UMAP of 60,000 cells normalized to sample size and proportional to the absolute cell number per group of Nestin^Cre/+Il12rb2^fl/fl (n = 11, m/f), Il12rb2^fl/fl (n = 7, m/f), Il12rb2^del/del (n = 9, m/f) at peak EAE (13 d.p.i.). f, Heatmap depicting median marker expression per cluster. g, Cell counts of CNS CD4⁺ T cells and MdCs corresponding to e. Data were pooled from two experiments (unpaired two-tailed t-test; *P = 0.03681; **P = 0.00203; *P = 0.02085; **P = 0.00151, left to right). h, Dot plot depicting fold change expression and P value of the indicated markers per population. i, Cohen’s d effect size for the expression of the indicated activation molecules in microglia from Nestin^Cre/+Il12rb2^fl/fl and Il12rb2^fl/fl mice. j, Representative WM staining with FluoroMyelin Green dye (top, thoracic; bottom, lumbar) and the percentage of myelin loss quantification in axial spinal cord sections at 19 d.p.i. in Nestin^Cre/+Il12rb2^fl/fl and control mice (n = 4 mice per group). Insets highlight active, demyelinating inflammatory lesions with accumulation of DAPI⁺Iba1⁺ phagocytes. Individual lesions are pointed out by arrowheads. Scale bars, 50 μm and 200 μm. Unpaired two-tailed t-test, ****P < 0.0001). k, Kinetics of IL-12 (IL-12/23p40) concentration (ELISA) in the brain and spinal cord in C57BL/6 mice n = 2–5 per time point in EAE, m/f; number of dots represents the number of biologically independent replicates per sample. l,m, Il12a (l) and Il12b (m) mRNA expression in FACS-isolated CNS CD45⁺CD44⁺CD11b⁺Ly6G⁻Ly6C⁺ MdCs (n = 7) and CD45^loCD44^loCD11b⁺CX3CR1⁺ microglia (n = 4) of C57BL/6 mice (m/f) at disease onset (10 d.p.i.). Unpaired two-tailed t-test, P = 0.053; *P = 0.0137, left to right. n,o, IL-12 protein in brain and spinal cord lysates (n) and clinical score of Nestin^Cre/+Il12rb2^fl/fl (n = 4, m) and littermate control mice (n = 4; m) at 10 d.p.i. (o). Two-tailed Mann–Whitney test, P = 0.1335; P = 0.5916; P = 0.7429; left to right). Each symbol represents one animal. Data are shown as the mean ± s.e.m.; m, male; f, female.

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