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. 2017 Jan 6;12(1):e0169621. doi: 10.1371/journal.pone.0169621

Fig 4. Aox sustains a membrane potential (ΔΨ) when coupled to complex I.

Fig 4

(A) Representative plot of mitochondrial membrane potential measurements in intact cells grown in glucose media Where indicated, 25 mM glucose, 0.5 μM antimycin A (AA), 3 mM SHAM and 10 μM CCCP (U) were added. Cells were incubated in regular respiration media at a final concentration of 1 mg.mL-1 (B) Aox-dependent ΔΨ measured as the difference between maximum fluorescence (glucose) and the fluorescence after Aox inhibition (SHAM). (C-E) Representative ΔΨ plots of isolated mitochondria from cells grown in YPD. (C) Complex I-dependent ΔΨ (10 mM pyruvate/malate/citrate each). Traces: Black, control without inhibitors. Red, Aox-sustained ΔΨ, in the presence of 100 μM cyanide (addition where indicated). Green/Blue, contribution of the cytochromes to the ΔΨ after the addition of 100μM salicylhydroxamic acid (SHAM, green) and 1 μM propyl gallate (PG, blue). (D) ΔΨ- dependent on Aox. Traces: Black, control without inhibitors. Blue/Red after addition of 100μM SHAM (blue) and 1 mM PG (Green) respectively. (F) Succinate/rotenone (10 mM/ 1 μM) dependent ΔΨ. Traces: Blue, complete collapse of ΔΨ after 100 μM cyanide addition. Black/Red, inhibition of Aox by SHAM (black) or PG (red) did not exert any effect on the ΔΨ, complete collapse of ΔΨ under these conditions was obtained after addition of 100 μM cyanide. At the end of all plots, 1 μM CCCP (U) was added to completely collapse the ΔΨ. * p < 0.05 respect to control cells.