Fig. 6.

Metabolic profiles of iTreg cells differentiated from naïve CD4⁺ CD25⁻ cells under different conditions. (A) Naïve CD4⁺ CD25⁻ cells pretreated with (closed symbols) or without (open symbols) Met were stimulated with antibodies against CD3 or CD3 and CD28 for three days in the presence or absence of TGF-β, as indicated. The OCR and ECAR were assayed using a Seahorse XF96 analyzer with the addition of anti-mitochondrial reagents, oligomycin, FCCP, rotenone (R), antimycin (A), and 2-deoxyglucose (2-DG). Representative data from three independent experiments are shown. (B) Basal levels of OCR, ECAR, and proton leakage are shown under each culture condition as in (A). The ECAR was assayed in Dulbecco's modified Eagle's medium without glutamate. (C) The ratio of ECAR/OCR was plotted as a bar graph. Notably, Met pretreatment decreased OCR/ECAR in iTreg. *P < 0.05; **P < 0.01.