Fig. 1.

Experimental design for the assessment of the effect of GRX2 deficiency on native ROS release rates from different sites of production. Oxidation of Krebs cycle linked metabolites, pyruvate and 2-oxoglutarate, allows the assessment of ROS release rates from pyruvate dehydrogenase (PDH) and 2-oxoglutarate (OGDH) and the electron transport chain. Succinate was used to test ROS release from the electron transport chain since it feeds electrons directly into the ubiquinone pool (Q pool), bypassing the Krebs cycle. The site of action for the selective ROS release inhibitors (KMV; 3-methyl-2-oxo valeric acid, rot; rotenone, A5; atpenin A5, myxo; myxothiazol), which allow the estimation of rates of ROS release production from individual sites, are indicated in the diagram. Sites for ROS production in the different experimental systems are denoted with a red star. Direction of electron flow is indicated by a dotted arrow.