(A) Enriched T cells (Teff) were labeled with CFSE, stimulated with soluble anti-CD3, and cultured alone or cocultured with iTregs at various ratios as indicated. The culture system was either in normal media or with indicated increased NaCl concentrations. Baseline indicates no added conditioned iTregs. CFSE dilution was detected by FACS after 3 days. Histograms depict cellular proliferation and are gated on CD8+ cells.
(B) The line graph depicts a summary of experiments at iTreg to Teff ratios as indicated (n = 5).
(C) Naive CD4 cells were differentiated into Tregs as described in STAR Methods, then re-stimulated under the indicated increased NaCl concentrations for another 3 days. Following incubation, iTregs were washed, counted and plated in a suppression assay in normal media as in (A). CFSE dilution was detected by FACS after 3 days. Histograms depict cellular proliferation and are gated on CD8+ cells.
(D) The line graph depicts a summary of experiments at iTreg to Teff ratios as indicated (n = 5).