TCF-1 controls Tc17 generation and MAF expression in developing thymocytes. (A) Flow-cytometric analyses and enumeration of RORγt and TCF-1 expression in TCRβ⁺CD8⁺CD62L⁻CD44⁺ splenocytes isolated from Tcf7^fl/flCD4Cre^+/+ and Tcf7^fl/flCD4Cre^+/T mice (upper panels) and Tcf7^fl/fldLckCre^+/+ and Tcf7^fl/fldLckCre^+/T mice (lower panels). Data are pooled from two independent experiments and show the mean ± SEM (n = 4 or 5 mice/genotype/experiment). Exact P values were calculated using Student’s t test. (B) Geometric MFI of MAF and RORγt expression in DP thymocytes (CD4⁺CD8⁺TCRβ⁻CD69⁻) and CD8 SP thymocytes (CD4⁻CD8⁺TCRβ⁺). Data are representative of two independent experiments (n = 4 mice/group/experiment) and show the mean MFI from one experiment. Exact P values were calculated using the Mann–Whitney U test. (C) Representative histograms of MAF and RORγt expression in DP thymocytes (CD4⁺CD8⁺TCRβ⁻CD69⁻) from B. (D) Genome browser of normalized TCF-1 ChIP-seq reads across the Maf locus is shown in green identified by ChIP-seq dataset from Dose et al. (2014) from WT mouse thymocytes. (E) Venn diagram analyses of up-regulated genes as determined by RNA-seq of Tcf7^−/− Tc17 cells versus Tcf7^+/+ effector cells, compared with TCF-1 target genes identified by ChIP-seq dataset from Dose et al. (2014). FDR of 10⁻⁵ was applied when calling ChIP-seq peaks. Selected shared TCF-1 target genes DE by Tc17 cells are shown with key transcription factors highlighted in red. (F) ChIP analysis of TCF-1 binding in Maf loci of mouse thymocytes. TCF-1 binding in the Axin2 loci is shown as a positive control. Data show the mean ± SD pooled from two independent experiments.