Fig. 3

SIRT5 interacted with c-myc to inhibit the succinylation of c-myc at K369 site. A, The succinylation level in sh-NC and sh-SIRT5 groups in HEK-293T cells was assessed by Western blot; B, STRING database was used to screen SIRT5-related proteins; C, Co-IP assay was performed to detect the interaction between SIRT5 and c-myc; D, The protein levels of c-myc and c-myc-succ in HEK-293T cells were detected by IP and Western blot after SIRT5 knockdown; E, The succinylation sites for c-myc were predicted using GPSuc database; F, IP and Western blot assays were used to analyze the succinylation sites of c-myc; G, The protein expression of c-myc was assessed by Western blot at the different time points (0, 8, 16, and 24 h) in HEK-293T cells; H, Quantification of the existing c-myc protein level at different time points (0, 8, 16, and 24 h) in HEK-293T cells. SIRT, sirtuin; STRING, Search Tool for Recurring Instances of Neighbouring Genes; Co-IP, co-immunoprecipitation; succ, succinylation