Fig. 1

Identification of Foxo1 in differentiated Th9 cells. Naive CD4⁺CD62L⁺CD44⁻ T cells were isolated from wild- type (Wt) C57/BL6 mice were and then differentiated into helper T (Th) 9 (TGF-β1 + IL-4) or Th0 (without skewing cytokines) conditions. Samples were collected for RNASeq analysis at 72 h time point. Unsupervised analysis of Th9 and Th0 cells transcriptome data, a PCA analysis and b Clustering analysis. Significantly differentially expressed genes between Th9 and Th0, c heat-map of all significantly differentially expressed genes, d heat-map analysis of selected top significantly differentially expressed genes. System biology analysis of the key transcriptional regulators that are activated e and inhibited f in Th9 Cells as compared to Th0 cells. g qPCR analysis of Il9 and Foxo1 in Th9 cells induced by TGF-β1 + IL-4, results were normalized to the expression of mouse Gapdh, and are presented relative to those of control Th0 cells. Bar shows mean ± s.d. from combined three experiments. ***P < 0.001. ****P < 0.0001 (unpaired student t-test). Flow cytometry analysis of total h and phospho Foxo1 i in Th0 and Th9 cells, data representative of two independent experiments