SIRT3 deacetylase activity assay

SIRT3 deacetylase activity was examined using a SIRT3 Fluorimetric Activity Assay Kit (#ab156067, Abcam) following the manufacturer's protocol. In brief, the mitochondrial extract was incubated with the SIRT3 assay buffer, and then co-incubated with Fluoro-Substrate Peptide, NAD and Developer at 37 °C for 1 h. Fluorescent intensity was measured using a Fluoroskan Ascent® microplate fluorometer (Thermo Electron Corp., Milford, MA, USA) at 350 nm/450 nm ⁷⁷.