Fig. 2: ABHD11 inhibition rewires mitochondrial metabolism in human T-cells.
From: Mitochondrial ABHD11 inhibition drives sterol metabolism to modulate T-cell effector function
a α-ketoglutarate dehydrogenase (α-KGDH) activity in CD4+ effector T-cells (n = 8). b Oxygen consumption rate (OCR) in CD4+ effector T-cells (n = 5). Pre-optimised injections include: oligomycin, FCCP, antimycin A/rotenone (all 1 μM) and monensin (20 μM). c Calculated OCR parameters from OCR measured in (b), including basal respiration, ATP-linked respiration, spare respiratory capacity and maximal respiratory capacity (n = 5). d Basal and maximal ATP production (JATP) from OCR measured in (b; n = 5). e Intracellular levels of selected TCA cycle intermediates in CD4+ effector T-cells. Metabolites include: acetyl-CoA, citrate/isocitrate, α-ketoglutarate (α-KG), succinate and malate (acetyl-CoA, citrate/isocitrate, α-KG, malate: n = 5; succinate: n = 4). f Determination of intracellular α-ketoglutarate to succinate ratio in CD4+ effector T-cells (n = 4). g Intracellular levels of selected amino acids in CD4+ effector T-cells. Metabolites include: glutamine, glutamate, aspartate and asparagine (glutamine: n = 4; glutamate, aspartate, asparagine: n = 5). h Mitochondrial content, as determined by MitoTracker™ Green, in CD4+ effector T-cells (n = 7). i Mitochondrial reactive oxygen species, as determined by MitoSOX™ Red, in CD4+ T-cells at 1 h (n = 5). j Extracellular acidification rate (ECAR) in CD4+ effector T-cells (n = 5). Pre-optimised injections include: oligomycin, FCCP, antimycin A/rotenone (all 1 μM) and monensin (20 μM). k Calculated ECAR parameters from ECAR measured in (j), including basal glycolysis and maximal glycolysis (n = 5). l Basal and maximal ATP production (JATP) from ECAR measured in (j; n = 5). m Intracellular levels of lactate in CD4+ effector T-cells (n = 5). n Extracellular lactate in cell-free supernatants from cultures of CD4+ effector T-cells (n = 15). All experiments were carried out using human samples. CD4+ T-cells were activated with α-CD3 (2 μg/ml) and α-CD28 (20 μg/ml) for 24 h, in the presence and absence of ML-226, unless otherwise stated. Data are expressed as either: mean, with paired dots representing data from distinct biological replicates; or mean ± SEM. Statistical tests used: two-tailed one-sample t-test (a), two-tailed paired t-test (c, e–i, k, m, n), two-way ANOVA with Šidák’s multiple comparisons test (d, l). Source data are provided as a Source Data file.
