Role of Setd2 in regulatory T cells [Pol II CUT&Tag]

To determine the molecular regulation of Tregs by Setd2, spleen Tregs (CD3+CD4+Foxp3-YFP+) from Setd2-deficient and control Tregs were subjected to polII CUT&Tag analysis IL-2/anti-IL-2 complexes (IL-2c) were prepared by mixing 1 ug recombinant mouse IL-2 (Peprotech) with 5 ug anti-IL-2 (bioxcell, clone JES6-1) and incubated at 37°C for 30 min. Foxp3Cre-YFPSetd2f/+ or Foxp3Cre-YFPSetd2f/f mice (n=2) were injected intraperitoneally (i.p.) with IL-2c for three consecutive days and sacrificed for analysis 48 h later. Splenic Tregs (CD3+CD4+Foxp3-YFP+) were sorted by flow cytometry and nuclei were extracted. Biological replicates of 80000 nuclei from each group were subjected to RNA polymerase II (Pol II ) CUT&Tag analysis.