Total RNA was isolated from CD4+ YFP(FOXP3)- T cells from pooled spleen and peripheral lymph nodes (cervical and inguinal) from Foxp3Cre WT mice and Foxp3-Cre Tet2/3fl/fl mice (14-weeks-old) using Direct-zol RNA microprep (Zymo, #R2060). RNA-sequencing libraries were prepared using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs, #E7760, #7765) according to the manufacture's protocol