FIG 4.

Acyl-K122 mimics of SOD1 inhibit SOD1-mediated suppression of mitochondrial respiration. (A) (Left) HEK 293 cells were transfected with a Flag-tagged WT, K122R, K122E, or K122Q SOD1 expression plasmid, and oxygen flux was measured with an Oroboros O2K respirometer. Cell lysates were separated by SDS-PAGE and immunoblotted for Flag-SOD1. The experiment was carried out in quadruplicate. Asterisks indicate significant differences (*, P < 0.05; **, P < 0.01); error bars represent SEM. (Right) HEK 293 cells were transfected with an empty vector (EV) or a Flag-tagged WT or G85R SOD1 expression plasmid and were assayed as in the experiment for which results are shown on the left. The experiment was carried out in triplicate. (B) HEK 293 cells were transfected with an empty vector or a Flag-tagged WT SOD1 expression vector. Oxygen rates were measured with an Oroboros O2K respirometer after inhibition of complex I (CI) with rotenone (2 mM) or of CII with malonate (2.5 mM). Three technical replicates were carried out.