Fig. 2

CD4⁺c‐Met⁺ T cells expressed higher level of Itgα4 and chemokine receptors in MOG_35-55-induced EAE at peak disease. A Heat map analysis of microarray data showing differentially expressed genes of CD4⁺c-Met⁻ and c-Met⁺ T cells isolated from CNS at EAE peak disease (d14) mice. The global gene expression profiles were analyzed by Principal Component Analysis (PCA). The first three principal components of microarray analysis data (PC1, PC2 and PC3) in X, Y and Z, respectively, are shown, and demonstrated the expression profile of three samples/groups (CD4⁺c-Met⁻ blue#1–3; CD4⁺c-Met⁺ Red#1–3). B Volcano plots comparing genes expressed by CD4⁺c-Met⁻ and c-Met⁺, showed a total of 1372 differentially regulated genes (1083 up-regulated, red, and 289 down-regulated, green) with at least twofold changes by P value (y axis) and fold change (x axis). C Heat map analysis of microarray data showing hierarchical clustering of differentially expressed probes between CD4⁺c-Met⁻ and CD4⁺c-Met⁺ cell population (three samples per group). Red and blue colors indicate differentially up- or down-regulated genes, respectively. The mean signals were background corrected and transformed to the log2 scale. Gene comparisons were done for CD4⁺c-Met⁺ and CD4⁺c-Met⁻ with at least twofold changes and p < 0.05 at the 95% confidence level were considered as significant. D KEGG pathways analysis revealed that selected genes were involved in several key pathways related to adhesion molecules associated with regulation of EAE development under an inflammatory microenvironment. Gene expression profiles are represented in fold change by CD4⁺Vα3.2⁺c-Met⁺ cells compared to CD4⁺Vα3.2⁺c-Met⁻ cells. E KEGG pathways analysis revealed that selected genes involved in Th1 (left panel) or Th17 (right panel) were upregulated in CD4⁺Vα3.2⁺c-Met⁺ cells when compared to CD4⁺Vα3.2⁺c-Met⁻ cells. Gene expression profiles are represented in fold change by CD4⁺Vα3.2⁺c-Met⁺ cells compared to CD4⁺Vα3.2⁺c-Met⁻ cells. F Representative flow cytometric histograms (left panels) and Gmean/isotype quantifications (right panels) of integrins α4 (Itgα4, VLA4 subunit) and αL (ItgαL, LFA1 subunit) expression by CD4⁺c-Met⁻ and c-Met⁺ T cells from spleen, LN and CNS at peak disease (d14). G Representative flow cytometric histograms (left panels) for chemokine receptors expression, such as CXCR3, CCR5 and CCR6 in CD4⁺c-Met⁻ and c-Met⁺ T cells isolated from spleen, LN and CNS at peak disease (d14). Gmean/isotype quantifications (right panels) representing mean values ± SEM for n = 5 mice/group are shown; *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001 by two-way ANOVA followed by Tukey’s post hoc test