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In vitro desuccinylation assay Hypersuccinylated GCLC proteins purified from HEK293T cells were incubated with purified SIRT2 in the presence or absence of 1 mmol/L NAD+ at 37 C for 4 h with desuccinylation buffer (50 mmol/L Tris-HCl [pH 7.5], 150 mmol/L NaCl, 1 mmol/L MgCl 2 )