Fig. 2

Nutrients promote ECHS1 K101 acetylation. a–c Western blot detection of acetylation levels of ectopically expressed ECHS1 in HEK293T cell cultured in the presence of excessive glucose (a), FAs (b), and amino acids (c) in the culture media. d Levels of endogenous ECHS1 in HEK293T and HepG2 cells were compared with and without the presence of deacetylases inhibitors NAM/TSA. e Effects of deacetylase inhibitors on the acetylation levels of affinity-purified endogenous ECHS1. f The MS spectrum that led to the identification of K101 acetylation containing tryptic peptide from endogenous ECHS1. g Western blot detection of proteins levels, AcK levels and AcK101 levels of endogenous ECHS1 in HEK293T cells that were cultured in DMEM base and DMEM base supplemented with glucose (25 mM), fatty acids (250 nM), and amino acids (8 mM). h Flag-tagged wildtype, K101R (K/R), and K101Q (K/Q) ECHS1 were overexpressed in HEK293T cells that were cultured in media with or without NAM/TSA. The relative acetylation levels of ECHS1 proteins purified from the above mentioned cells using Flag beads was determined. For b–e and h, mean values of quantitation with SD are reported. *P < 0.05; **P < 0.001; ***P < 0.001