Figure 1.

Gating Strategy for FACS Isolation of Paired CD4⁺CD25⁺CD127^lo/- and CD4⁺CD25⁺CD226^- Tregs. Representative flow plots demonstrate the method by which CD127^lo/- or CD226^- Tregs were isolated from CD4⁺ T-cell enriched PBMC using a BD FACSAriaIII Cell Sorter. (A) Singlet gating was performed using forward scatter area (FSC-A) versus forward scatter height (FSC-H). (B) Lymphocytes were gated on FSC-A and side scatter area (SSC-A). (C) From the CD4⁺ T cell fraction, (D) CD25⁺CD127^lo/- Tregs and (E) CD25⁺CD226^- Tregs were isolated.