Figure 1. ZEB1 mutations modulate risk for multiple sclerosis and experimental autoimmune encephalitis.

(A) Schematic to identify putative MS-relevant regulators of pathogenic Th1/Th17 differentiation.

(B) Heatmap of MS-associated genes expression in CD4⁺ T cells and monocytes from MS patients.

(C) Bar plot showing significantly enriched pathways in the genes in (B).

(D) Heatmap of genes highly expressed in Th1 and/or Th17 cells versus Th2 and Treg cells.

(E) Dot plot showing ZEB1 gene expression level in myelin-reactive Th17 cells from MS patients (Th17 MS) and healthy controls (Th17 Ctrl).

(F) Dot plot showing linear correlation of ZEB1 gene expression with IL-17A and IFNG gene expression in myelin-reactive Th17 cells from MS patients (Th17 MS) and healthy controls (Th17 Ctrl).

(G) Schematic displaying the experimental design for experimental autoimmune encephalomyelitis (EAE) model.

(H) Clinical scores of wild-type (WT, n = 11) and CD4^CreZEB1^L/L (n = 7) mice in EAE. Statistic difference is tested by nonparametric Mann-Whitney test.

(I) Flow cytometry evaluating the number of CD4⁺ cells, CD4⁺IL-17A⁺ cells, CD4⁺IFN-γ⁺ cells, CD4⁺GM-CSF⁺cells, and CD8⁺ cells in the CNS of CD4^CreZEB1^L/L (n = 4) and WT (n = 5) mice at 18 days post immunization.

(J) Flow cytometry evaluating the number of CD4⁺ cells, CD4⁺IL-17A⁺ cells, CD4⁺IFN-γ⁺ cells, CD4⁺GM-CSF⁺cells, CD4⁺CD25⁺FOXP3⁺ cells, and CD8⁺ cells in the spleen of CD4^CreZEB1^L/L (n = 4) and WT (n = 5) mice at 10 days post immunization.

(K) Flow cytometry analysis of cytokine expression in CD4⁺ T cells from reactivated splenocytes from CD4^CreZEB1^L/L and WT mice.

The data in (E) and (H)–(K) are presented as the mean ± SEM.

Statistical differences in (F) were tested using Pearson’s correlation coefficient test. All statistical differences in (E) and (I)–(K) were tested using unpaired Student’s t test (two-tailed). *p < 0.05, **p < 0.01.