Given the reduced percentage of CD25int mTreg that possess a demethylated TSDR after expansion compared with naive Treg cells (Figure 3A), it is possible that CD25int mTreg may have the potential to revert to an effector function. To test whether the reduced TSDR demethylation corresponds with enhanced effector function, we examined IFN-γ and IL-17A production after expansion. Indeed, CD25int mTreg produce substantially greater levels of both IFN-γ and IL-17A compared with naive Treg (Figure 5). Tacrolimus significantly reduced both IL-17A and IFN-γ production (Figure 5).