Fig. 4: Mitochondrial metabolism differences between human and mouse astrocytes.

a Basal oxygen consumption rate (OCR) of mouse and human astrocytes. Each data point (circle or square) represents one well of astrocyte culture prepared from one human patient or one litter of 8–10 mice throughout this figure. Mouse astrocytes: N = 7 cultures generated from 4 litters of mice in a–c. Human astrocytes: N = 6 wells of cultured cells generated from 3 patients in a–c. p = 0.0365. Two-tailed unpaired Welch’s t-test. The p-values were calculated using average results from each litter of mice and each patient as independent observations, unless otherwise indicated. b OCR linked to ATP production in the presence of oligomycin. p = 0.0497. Two-tailed unpaired Welch’s t-test. c The ratio of OCR to extracellular acidification rate (ECAR). p = 0.0168. Two-tailed unpaired Welch’s t-test. d, e Tetramethylrhodamine, ethyl ester (TMRE) fluorescence (reporting mitochondrial membrane potential) normalized by MitoTracker Green (MTG, a general mitochondrial dye) fluorescence. Data represent H₂O₂-treated conditions normalized to medium control-treated conditions. N = 9 wells of cultured cells generated from 3 litters of mice and 4 patients. Mouse astrocytes: 3 hr vs. 1 hr, p = 0.3959. Human astrocytes: 3 hr vs. 1 hr, p = 0.0373. Two-tailed unpaired Welch’s t-test. N.S., not significant. f, g Basal OCR of astrocytes treated with 100 μM H₂O₂. 0 hr and 1 hr mouse astrocytes: N = 7 wells of cultured cells generated from 4 litters of mice. 3 hr and 5 hr mouse astrocytes: N = 8 wells of cultured cells generated from 4 litters of mice. 0 hr, 3 hr, and 5 hr human astrocytes: N = 6 wells of cultured cells generated from 3 patients. 1 hr human astrocytes: N = 5 wells of cultured cells generated from 3 patients. Mouse astrocytes: 0 hr vs. 1 hr, p = 0.1883; 0 hr vs. 3 hr, p = 0.4246; 0 hr vs. 5 hr, p = 0.0285. Human astrocytes: 0 hr vs. 1 hr, p = 0.0954; 0 hr vs. 3 hr, p = 0.0235; 0 hr vs. 5 hr, p = 0.1837. One-way repeated measure ANOVA with Dunnett’s multiple comparison test. h, i ATP-linked OCR of astrocytes treated with 100 μM H₂O₂. The replicate numbers are defined in (f–g). Mouse astrocytes: 0 hr vs. 1 hr, p = 0.8885; 0 hr vs. 3 hr, p = 0.7597; 0 hr vs. 5 hr, p = 0.2198. Human astrocytes: 0 hr vs. 1 hr, p = 0.1639; 0 hr vs. 3 hr, p = 0.0016; 0 hr vs. 5 hr, p = 0.2583. One-way repeated measure ANOVA with Dunnett’s multiple comparison test. j Diagram of glucose metabolism. TCA, tricarboxylic acid. NADPH, nicotinamide adenine dinucleotide phosphate.