Oxytherm r system

Oxygen consumption was performed as described previously [21 (link)]. Briefly, cells were collected at their logarithmic phase and 2 × 10⁷ cells were resuspended in 1 mL of YE5S to a final OD₆₀₀ of 1. The measurements were made using Oxytherm + R system (Hansatech, Pentney, UK), with the standard setting to measure in nmoles/mL, recorded over 10 min. The addition of carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) (6 µM) induced maximal respiration by uncoupling electron transport from oxidative phosphorylation. Each one of the measurements was performed from biological triplicates.

Oxygen consumption was measured using an Oxytherm+R system (Hansatech instruments, Norfolk, UK)^{12 (link)}. The system was calibrated daily with air saturated deionized water (high O₂) and glucose oxidase + glucose. 10⁶ cells were washed in PBS and permeabilized in solution containing (in mM) 125 KCl, 10 K₂HPO₄, 1 MgCl₂, 0.01 EGTA, 20 HEPES, pH 7.2 supplemented with 50 µg/mL digitonin, for 15 min on ice. Oxygen consumption was monitored after the serial addition of 5 mM L-glutamic acid + 5 mM L-malic acid, 0.5 mM ADP, and 5 mM succinic acid + 1 µM rotenone.