Figure 4.

Mito‐transfer increases aerobic metabolism in CD4⁺ T cells from old mice. CD4⁺ T cells (2.5 x 10⁵) were plated onto poly‐D‐lysine coated 96‐well microplates, after which O2 consumption (OCR) and extracellular acidification rates (ECAR) were measured under basal conditions and in response to the mito‐stress test. A) Representative mito stress test kinetic graph, B) quantitation of basal, maximal, and spare respiratory capacity, C) proton leak, non‐ mitochondrial oxygen consumption, and ATP production rate, and D) coupling efficiency. E) Representative glycolysis stress test kinetic graph, and quantitations of F) non‐glycolytic acidification, glycolysis, glycolytic capacity, glycolytic reserve and G) GlycoPER of CD4⁺ T cells. H) Flow cytometry histograms and I,J) dot plots of 2‐NBDG uptake in CD4⁺ T cells from young and old mice, and CD4⁺ T cells from old mice after mito‐ transfer. K) Flow cytometry histograms and L) dot plots of GLUT1 expression in CD4⁺ T cells from young and old mice, and CD4⁺ T cells from old mice after mito‐transfer. M) Rates of glycolytic and mitochondrial‐derived ATP, and total ATP production, and N) cellular levels of ATP in CD4⁺ T cells from young and old mice, and CD4⁺ T cells from old mice after mito‐transfer; 3 to 4 mice were used per group and experiments were repeated at least once (n ≥ 2), p ≤ 0.05 = *, p ≤ 0.01 = **, p ≤ 0.001 = *** and p ≤ 0.001 = ****, using one‐way‐ANOVA.