Cell-based and multi-omics profiling reveals dynamic metabolic repurposing of mitochondria to drive developmental progression of Trypanosoma brucei
Fig 5
RBP6OE cells respire predominantly via AOX.
(A) The resting respiration of cells undergoing RBP6-induced differentiation was measured using the O2k-oxygraph. The ratio of complex IV–and AOX-mediated respiration was determined using KCN, a potent inhibitor of complex IV, and SHAM, a potent inhibitor of AOX. Individual values shown as dots (mean ± SD, n = 3–5), ****P < 0.0001. (B) Glycerol-3-phosphate stimulated respiration in live cells. The proportion of complex IV–and AOX-mediated respiration was determined as in (A). Individual values shown as dots (mean ± SD, n = 5–8), **P < 0.01. (C) In vitro ATP production was measured in digitonin-extracted mitochondria. The OxPhos pathway was triggered by the addition of ADP and glycerol-3-P. Treatment with 1 mM KCN serves as a control. Individual values shown as dots (mean ± SD, n = 3–4). Underlying data plotted in panels (A), (B), and (C) are provided in S1 Data. AOX, alternative oxidase; Gly-3-P, glycerol-3-phosphate; KCN, potassium cyanide; OxPhos, oxidative phosphorylation; RBP6, RNA binding protein 6; SHAM, salicylhydroxamic acid.