Fig. 3

CHD8-deficient Tregs become plastic and are outcompeted by CHD8-proficient Tregs. A–D Freshly isolated splenocytes were stimulated with PMA plus ionomycin for 5 h with BD GolgiPlug™ added during the last 2 h. The cells were collected for surface staining of CD4 and intracellular staining of Foxp3, IFN-γ IL-4, IL-13, and IL-17. Percentages of IFN-γ⁺ (A), IL-4⁺ B, IL-13⁺ (C), and IL-17⁺ (D) cells gated on CD4⁺Foxp3⁺ cells are shown in representative dot plots and summarized in bar graphs. E Percentages of YFP⁺ and YFP⁻ cells gated on CD4⁺Foxp3⁺ cells are shown in representative dot plots from Chd8^+/+Foxp3^YFP−cre/+ and Chd8^Flox/FloxFoxp3^YFP−cre/+ mice. The ratios of YFP⁺/YFP⁻ cells are summarized in a bar graph. F Expression of the indicated Treg functional markers on Foxp3⁺YFP⁻ and Foxp3⁺YFP⁺ cells from Chd8^Flox/FloxFoxp3^YFP−cre/+ mice. Numbers indicate % and MFI. Data are representative of five mice. Results in bar graphs represent mean plus SD (WT: three males + three females; Chd8^−/−: two males + three females). **P < 0.01 versus WT control group