FIG. 4.

aTregs from BDC12-4.1.RAG^KO mice coexpress IFN-γ and IL-10, whereas Teffs produce solely IFN-γ. Total splenocytes from BDC12-4.1.RAG^KO (A and B) and BDC12-4.1.RAG⁺ (C and D) were incubated in vitro for 4 h with BfA (10 μg/mL) either alone (control, no activation) or in the presence of BfA and anti-CD3/28 (10/5 μg/mL, respectively) or phorbol myristic acid/ionomycin (50/1000 ng/ml, respectively). Cells were costained for Foxp3, IFN-γ, and IL-10 after fixation with the Foxp3 Fix/Perm kit. The cytokine profile is shown after gating on the Treg (CD4⁺Foxp3⁺) or non-Treg (CD4⁺Foxp3⁻) population. In the control (no activation), gating was performed on total CD4⁺ cells. One representative mouse (for each genotype) of at least three that were tested in two independent experiments is shown. (A high-quality color representation of this figure is available in the online issue.)