Extended Data Fig. 1: 13C3-Pyruvate and malate feeding to isolated mitochondria of Col-0, me1.me2 and me1.me2.mpc1.
From: Metabolic evidence for distinct pyruvate pools inside plant mitochondria
(a) Time courses of metabolite concentrations in the extra-mitochondrial space of isolated mitochondria from Col-0 incubated with various concentrations of 13C3-pyruvate and 500 µM malate (n = 3). (b) Time courses of metabolite concentrations in the extra-mitochondrial space of isolated mitochondria from Col-0, me1.me2, me1.me2.mpc1 incubated with 500 µM 13C3-pyruvate and 500 µM malate (n = 4). All experiments were conducted in the presence of ADP at pH 6.4 to initiate substrate uptake and consumption by both pathways - the MPC pathway (labelled metabolites) and the NAD-ME pathway (unlabeled metabolites). Metabolic reaction was stopped by centrifugation through a single silicon oil layer by which the mitochondrial pellet was separated from the extra-mitochondrial medium. Unused substrate and exported products in the extra-mitochondrial medium were quantified using LC-SRM-MS. Each data point represents averaged value from three or more biological replicates with error bars indicating standard error. Significant differences between mutants and wildtype are denoted by coloured asterisks based on two-sided Student’s t-tests (*, p < 0.05, See Source data Extended data Fig. 1 for exact p-values).
