Reversal of aPC’s effect on mitochondrial metabolism in CD4⁺CD25⁻ cells by α-KG or glutamine supplementation. (A) Line graph showing Seahorse analysis of OCR in human CD4⁺CD25⁻ cells preincubated without (T) and with aPC alone (T + aPC; 1 hour, 20 nM) or with supplementation of α-KG (48 hour, 3.5 mM, T + aPC + α-KG) or glutamine (48 hours, 4 mM, T + aPC + Q). (B) Schematic line plot showing the areas of the OCR curves representing basal respiration, maximum respiration, spare respiratory capacity (SRC), and adenosine triphosphate (ATP) production. (C-F) Bar graphs with dot plots summarizing basal respiration (C), maximal respiration (D), SRC (E), and ATP production (F) (n = 5) in experimental groups as described in panel A. The data are shown as the mean ± SEM; statistical significance was determined by 2-way analysis of variance (ANOVA) for panel A or 1-way ANOVA for panels C-F. Significance is represented in panel A: ∗P < .05 (T vs T + aPC), ^#P < .05 (T + aPC vs T + aPC + α-KG), ^$P < .05 (T vs T + aPC + Q); and panels C-F: ∗P < .05, ∗∗P < .01, and ∗∗∗P < .005.