Fig. 3. Mitochondrial copper(ii) regulates NAD(H) redox cycling.

a, Fluorescence microscopy of SOD2 in MDMs. Representative of n = 4 donors. At least 50 cells were quantified per donor. Scale bar, 10 μm. Two-sided unpaired t-test. Data are mean ± s.d. b, Representative western blots of SOD2 and catalase in MDMs (n = 7 donors). c, Regulation of H₂O₂ levels by SOD2 and catalase. d, Flow cytometry of mitochondrial H₂O₂ in MDMs (n = 6 donors). e, Reaction of NADH with H₂O₂ under copper(ii)-catalysed or copper-free conditions. Experimental mass spectrometry peaks and calculated masses of molecular ions are indicated. ES+, electrospray ionization mass spectrometry. f, Kinetics of NADH oxidation in the presence of H₂O₂ and copper(ii). Data are representative of n = 3 independent experiments. g, Metabolomics of NAD⁺ and NADH of mitochondria from MDMs treated with LCC-12 (n = 9 donors). AreaCorrLog₂Cen data correspond to raw areas, corrected for analytical bias using GRMeta R package, then corrected areas are log₂ transformed and centered on means. h, Metabolomics heat map highlighting metabolites whose biosynthesis is dependent on NAD(H) in MDMs treated with LCC-12 (n = 9 donors). i, Metabolomics of αKG and acetyl-CoA of MDMs treated with LCC-12 (n = 9 donors). b,d, Two-sided Mann–Whitney test. g,i, Kruskal–Wallis test with Dunn’s post test. In graphs, each coloured dot represents an individual donor for a given panel.

Source data