Fig. 2: LARS1 is O-GlcNAcylated at S1042 by OGT1 under glucose starvation.

a SW620 cells were incubated with or without 11 mM glucose for 4 h. b SW620 cells were transfected with the indicated siRNAs. After 48 h, the cells were starved of glucose for 4 h, then permeabilized with streptolysin O for 5 min, and 200 μM of UDP-GlcNAc, as indicated, was added for 30 min. c SW620 cells were starved of glucose and supplemented with 11 mM glucose for the indicated durations. d SW620 cells were incubated with vehicle or 20 μM ST045849 for 24 h and starved of glucose for 4 h. a–d Each cell lysate was precipitated with succinylated wheat germ agglutinin (sWGA)-conjugated agarose beads or anti-LARS1 antibody-conjugated agarose beads and then immunoblotted with the indicated antibodies. e The site of LARS1 O-GlcNAcylation was mapped using mass spectrometry. f LARS1 protein sequences, including S1042, the O-GlcNAcylation site, among species were aligned using BLAST. Red alphabet indicates a conserved serine residue. g SW620 cells were transfected with the indicated LARS1 constructs. After 24 h, the cells were starved of glucose for 4 h. Each sample was precipitated with sWGA-conjugated agarose beads or anti-LARS1 antibody-conjugated agarose beads and analyzed by immunoblotting with the indicated antibodies. Representative data of three experiments with similar results. Source data are provided as a Source Data file.