American Society of Anesthesiologists

Fig. 1.

Antimycin A and Pam3Cys induce intracellular H2O2 and mitochondrial O2- production in peritoneal leukocytes. (A) Antimycin A leads to a dose-dependent mitochondrial O2- production. Thioglycollate-elicited peritoneal leukocytes were treated with antimycin A for 1 h and analyzed for mitochondrial O2- production with flow cytometry. n = 3 in each group. *P < 0.05, **P < 0.01 vs. the untreated controls. Each error bar represents mean ± SEM. MFI, mean fluorescence intensity. The experiments were performed twice with similar results. (B, C) Antimycin A- or Pam3Cys-induced intracellular or mitochondrial reactive oxygen species production. Representative histograms of flow cytometry (B) and fluorescent images (C) are presented. Peritoneal leukocytes were treated with antimycin A (10 μg/ml) or Pam3Cys (20 μg/ml) for 1 h, incubated with 10 μM DCF or 2.5μM MitoSOX, and then analyzed for cellular H2O2 or mitochondrial O2- production, respectively, with flow cytometry (B) or fluorescent microscope (C). DCF = dichlorodihydrofluorescein diacetate; H2O2 = hydrogen peroxide; Mito O2- = mitochondrial superoxide; MitoSOX fluo = MitoSOX fluorescence.

Antimycin A and Pam3Cys induce intracellular H₂O₂ and mitochondrial O₂^- production in peritoneal leukocytes. (A) Antimycin A leads to a dose-dependent mitochondrial O₂^- production. Thioglycollate-elicited peritoneal leukocytes were treated with antimycin A for 1 h and analyzed for mitochondrial O₂^- production with flow cytometry. n = 3 in each group. *P < 0.05, **P < 0.01 vs. the untreated controls. Each error bar represents mean ± SEM. MFI, mean fluorescence intensity. The experiments were performed twice with similar results. (B, C) Antimycin A- or Pam3Cys-induced intracellular or mitochondrial reactive oxygen species production. Representative histograms of flow cytometry (B) and fluorescent images (C) are presented. Peritoneal leukocytes were treated with antimycin A (10 μg/ml) or Pam3Cys (20 μg/ml) for 1 h, incubated with 10 μM DCF or 2.5μM MitoSOX, and then analyzed for cellular H₂O₂ or mitochondrial O₂^- production, respectively, with flow cytometry (B) or fluorescent microscope (C). DCF = dichlorodihydrofluorescein diacetate; H₂O₂ = hydrogen peroxide; Mito O₂^- = mitochondrial superoxide; MitoSOX fluo = MitoSOX fluorescence.

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