Figure 4. Pathway analysis shows upregulation of inflammatory phenotype and mTORC1 signaling in migrated Tregs.

Bulk RNA-Seq was performed on untouched, migrated, and nonmigrated Tregs. (A and B) Volcano plot of DEGs in migrated versus nonmigrated Tregs for HD (A) and uRRMS (B). The x axis shows the log₂ fold change for ratio migrated/nonmigrated. The y axis shows statistical significance (FDR-adjusted P value). Up- and downregulated genes are colored red or blue, respectively, if adjusted P < 0.05 and |log₂ fold change| > 1. Inflammation-, migration-, and regulation-related genes are highlighted. (C) The 10 most significant DEGs were grouped into following pathways (GSEA identified; Supplemental Table 2, and Supplemental Figure 5): inflammatory response, TNF-α signaling, mTORC1 signaling, IFN-γ response, and IL-6-STAT3 signaling (only enriched in migrated uRRMS-derived Tregs). Relative gene expression is indicated by color: upregulation in red and downregulation in blue. Expression values are given as Z scores; Benjamini-Hochberg adjusted P value (FDR < 0.05) and |log₂ fold change| > 1; nonsignificant differences (FDR > 0.05 or |log₂ fold change| > 1) are marked with #. n = 5 (HD) and n = 4 (uRRMS)