Figure 1

Effect of carbon monoxide concentration in mitochondrial swelling. Mitochondrial swelling was followed by the decrease in absorbance at 540 nm at 37°C for 30 minutes. A. Mitochondrial swelling was assessed in the presence of CO at 0, 10, 25, 50, 100, 250 and 500 μM. Left panel, representative graphics of experiments, performed in triplicates and repeated, at least, three times. Right panel, quantitative expression of mitochondrial swelling measured at 28 minutes of incubation. All values are mean ± SD (error bars), n = 3; *p < 0.05, compared to control mitochondria. B. Rhodamine 123 fluorescence change (λ_exc: 485 nm, λ_em: 535 nm) was used to measure mitochondrial depolarization and to test CO toxicity in isolated liver mitochondria. Quantitative expression of mitochondrial depolarization measured at 15 minutes after CO treatment, all values are mean ± SD (error bars), n = 3; *p < 0.05, compared to non-treated mitochondria. C. Mitochondria were pre-treated with CO at 0, 10, 50 or 100 μM for 15 minutes at RT, and measurements were acquired at 37°C after Ca²⁺ at 12.5 μM addition to trigger swelling. Left panel, r epresentative graphics of experiments, which were done in triplicates and repeated, at least, three times. Right panel, quantitative expression of mitochondrial swelling measured at 12 minutes of incubation. The effect of 12.5 μM of Ca²⁺ was normalized as 100% of swelling. All values are mean ± SD (error bars), n = 3; *p < 0.05, compared to control mitochondria; **p < 0.05, compared to Ca²⁺ 12.5 μM treated mitochondria.