AMPK KO T cells reduce both oxidative and glycolytic metabolism. A total of 2 × 106 CD45.1+ WT (fl/fl) or AMPK KO (blue) T cells and 5 × 106 T-cell depleted (TCD) B6 bone marrow cells were transplanted into irradiated allogeneic (B6D2F1) recipients. On day 7 after transplant, donor T cells were purified by negative selection over a magnetic column, placed into the Seahorse metabolic analyzer, and metabolism interrogated using the mitochondrial stress kit. OCRs (A), including both maximal OCR (B) and SRC (C), were measured simultaneously with ECARs, as shown in panel D. Response to oligomycin (w/oligo) was calculated by subtracting individual values from the averaged baseline values prior to oligomycin administration (E). Maximal ECAR values were simply the highest ECAR values obtained over the course of the analysis (F). (G-L) Donor T cells were recovered from a separate cohort of B6D2F1 recipients on day 21 posttransplant and measured for OCR (G-H), SRC (I), ECAR (J), response to oligomycin (K), and maximal ECAR values (L). Panels A-F, n = 2 pooled samples per group (3-4 mice in each pool), with plots representative of 2 independent experiments. In panels G-L, n = 3 pooled samples, with 3 to 4 mice per pool. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001. fl/fl, floxed/floxed.
