Figure 4.

Acetylation/deacetylation status in MnSOD-K68 regulates mitochondrial metabolism. (a) One million of SOD^-/- MEFs overexpressing MnSOD^WT, MnSOD^K68R, and MnSOD^K68Q were stained with 100nM MitoTracker Green (Cell Signaling) solution diluted in 1X PBS and analyzed using FITC channel via flow cytometry. Stained cell populations were further analyzed using FlowJo; (b) SOD^-/- MEFs overexpressing MnSOD^WT, MnSOD^K68R, and MnSOD^K68Q were measured for ATP turnover rate determined by the difference between oxygen consumption rate (OCR) before and after adding ATP synthesis inhibitor oligomycin; (c) Basal respiration OCR was measured by the difference between OCR before and after adding complex I/III inhibitor rotenone/antimycin mixture; (d) Maximal respiration OCR was determined by the difference between OCR before and after adding mitochondria uncoupler CCCP/FCCP; (e) Proton leak rate was measured by the difference between the OCR after adding oligomycin and after adding rotenone/antimycin. All experiments were done in triplicates. Error bar represents ±1 SEM. *p<0.05, and **p<0.01. Three groups were analyzed using a one-way ANOVA test via GraphPad Prism 6.