Fig 4. GA-activated Ahr alleviates EAE severity.

The mice were immunized with MOG_35-55 emulsified in CFA. The mRNA expression of Cyp1a1 and miR-212/132 cluster were assessed in isolated CD4⁺ T cells and CD11b⁺ cells by quantitative real-time PCR and normalized to the mRNAs of Gapdh for Cyp1a1 and RNU6B for miRNAs. The levels of cytokines were quantified by ELISA. (A) EAE clinical score, maximum score and incidence (%) of Control EAE (Control) and GA EAE mice, n = 10 each. (B) Serum levels of IL-6, IL-1β and TNF-α in EAE mice 24 days after MOG_35-55 immunization. (C) Levels of IL-17, TGF-β and IL-10 in culture supernatant of encephalitogenic CD4⁺ T cells restimulated with MOG_35-55 and IL-23 for 72 h. (D) Frequency (%) of CD4⁺IL-17⁺ and CD4⁺Foxp3⁺ T cells in total CD4⁺ T cells isolated from inguinal lymph nodes 24 days after MOG_35-55 immunization. (E) Absolute number of CD4⁺CD45⁺ T cells in the CNS 18 days after MOG_35-55 immunization. (F) Relative expression of Cyp1a1 mRNA in CD4⁺ T cells and CD11b⁺ cells isolated from spleen 10 days after MOG_35-55 immunization compared to Control. (G) Relative expression of miR-132 and miR-212 in CD4⁺ T cells and CD11b⁺ cells isolated from spleen 10 days after MOG_35-55 immunization compared to Control. (H) Representative immunoblot of AChE in spleen WBCs 10 days after MOG_35-55 immunization. Data were pooled from three independent experiments and shown as mean ± SD. *p<0.05; (A) EAE score, two‐way ANOVA; (A) incidence (%), X²; (A) maximum score and (B-G), one-way-ANOVA; horizontal bars denote statistical comparison.