FIGURE 6.

Effect of H₂O₂ and diamide on mitochondrial proton leak. a, impact of successive H₂O₂ and diamide treatments on proton leak in mitochondria isolated from muscle of WT and Grx2^−/− mice. One-way analysis of variance with Fisher's protected least significant difference post hoc test, n = 4, mean ± S.E. * and # correspond to comparisons with WT or Grx2^−/− (oligomycin (oligo) conditions). b, impact of successive H₂O₂ (Per) and diamide (Dia) treatments on proton leak in mitochondria isolated from muscle of WT and UCP3^−/− mice. One-way analysis of variance with Fisher's protected least significant difference post hoc test, n = 4, mean ± S.E. * and # correspond with comparisons to WT or UCP3^−/− (oligomycin conditions). c, Grx2 is required to glutathionylate UCP3. Mitochondria were treated with BioGEE, and proteins were immunoprecipitated and then tested for UCP3 by immunoblot. d, visualization of UCP3 glutathionylation status by immunofluorescent microscopy. Gastrocnemius muscle sections were fixed and treated with antibodies directed against UCP3 (green) and BioGEE (red). Co-localization of both stains provides a yellow fluorescence. Preincubation in NEM or DTT were used as controls. Scale bar, 10 μm.