Figure 5
From: Epithelial cell-derived cytokine TSLP activates regulatory T cells by enhancing fatty acid uptake
TSLP-stimulated in vitro-induced Treg cells are more dependent on fatty acids. (A) Differentially expressed genes of TSLP-stimulated Treg cells. RNA-seq of PBS- or TSLP-stimulated Treg cells were analyzed. Genes significantly upregulated by TSLP are in red and downregulated are in blue. Symbols of top 20 genes are also labeled. n = 2 for each condition. (B) Enriched KEGG pathway of TSLP-stimulated Treg cells. Pathways with P-values less than 0.1 were depicted. Pathways with P-value less than 0.05 were marked with *. (C) Phosphorylation of S6 by TSLP. Left, representative histograms. Right, cumulative data of geometric mean fluorescence intensity (gMFI). n = 7 from 4 independent experiments. *P < 0.05 as determined by paired t-test. (D,E) Heatmaps of fatty acid transport/oxidation-related genes (D) and of glucose transport-related genes (E) in TSLP-stimulated and PBS-stimulated Treg cells. (F) Fatty acid uptake by TSLP. Naive CD4+ T cells were cultured under iTreg conditions in the presence or absence of TSLP for 3 days. Cells were pulsed with Bodipy FL C16 for 1 h and subjected to flow cytometry. Representative histograms and cumulative data are shown. n = 6 from 2 independent experiments. (G–H) Treg cell activation by TSLP in a fatty acid-free medium. Naïve CD4+ T cells were cultured under iTreg conditions in the presence or absence of TSLP for 3 days in RPMI medium supplemented with regular FBS or charcoal dextran-treated FBS. Cell size (G) and CD44 expression (H) of Treg cells were analyzed by flow cytometry. Representative FACS plots (for FSC and CD44) and cumulative data (for CD44) are shown. n = 5 from 2 independent experiments. *P < 0.05 as determined by unpaired t-test.
