Fig. 6

CPT1A binds to and succinylates LDHA at K222. a and b KAT2A does not bind to LDHA or its mutant K222E. AGS cells were transfected with Flag-KAT2A in combination with HA-LDHA or HA-K222E plasmid. KAT2A fusion protein was pulled down by Flag antibody, LDHA or K222E was detected using HA antibody. c-e CPT1A interacts with LDHA and its mutant K222E in AGS and 293 T cells. Cells were transfected with Flag-CPT1A in combination with HA-LDHA (c, d) or HA-K222E (e). The co-IP assay was performed similarly as above (a, b). f and g CPT1A promotes the succinylation of LDHA (f), but not of its mutant K222E (g). AGS cells were transfected with the indicated plasmids. Then, HA-LDHA was immunoprecipitated and its succinylation analyzed using either pan-succinylation or K222suc-specific antibody. h and i Knockdown efficiency of three CPT1A shRNAs in AGS cells was determined by western blot (h). CPT1A deficiency decreased the succinylation of LDHA (i)