Supplementary Figure 3: MSC-deficient iT_reg cells exhibit enhanced expression of T_H2-cell-related genes.

(a) The level of phosphorylated Smad3 was assessed by immunoblotting in naive T cells from WT and Msc^–/– mice stimulated with TGF-β for the indicated time; (b) Naive CD4⁺ T cells from WT and Msc^–/– mice were stimulated under iT_reg conditions with TGF-β and harvested at 72 hours. Relative mRNA expression levels of indicated genes are shown by q-PCR; (c) T_H2 RPKM values were than compared to all other T cell subsets (T_H1, T_H2, T_H17, T_H9 and T_reg, GSE 39756). Differentially expressed genes (fold change >1.5) in each comparison were than overlapped with genes that were upregulated in the Msc^–/– iT_reg cells (fold change >1.5). Enrichment was calculated as the number of overlapping genes divided by the total number of genes upregulated in each T cell subset. Data are pooled from three independent experiments (b) or are representative of three independent experiments (a).