Fig. 5: The O-GlcNAcylation of LARS1 regulates ULK1-mediated LARS1 phosphorylation.

a SW620 cells were starved and stimulated with 11 mM glucose for the indicated durations. b Quantification of LARS1 S720 phosphorylation and O-GlcNAcylation of a (mean ± SEM, n = 3 independent experiments). c, d SW620 cells were transfected with the indicated siRNAs. After 48 h, the cells were starved of glucose for 4 h, pre-incubated with streptolysin O (SLO) for 5 min, and supplemented with 11 mM glucose or 200μM indicated metabolites for 30 min. e SW620 cells were transfected with the indicated siRNAs. After 48 h, the cells were starved of glucose for 4 h with vehicle or indicated compounds (10μM Compound C, 20 μM SBI-0206965, 1 μM LYN1604, 1 mM AICAR) f SW620 cells were transfected with the indicated LARS1 constructs. After 24 h, the cells were starved of glucose for 4 h and supplemented with 11 mM glucose for 30 min. g SW620 cells were transfected with control or siRNA targeting OGT1. After 48 h, the cells were starved of glucose at the indicated durations. h SW620 cells were transfected with siRNA targeting ULK1. After 24 h, the cells were transfected with the indicated expression construct. After 24 h, the cells were starved of glucose for 4 h. Each sample was analyzed by immunoblotting with the indicated antibodies. i SW620 control or S1042A knock-in cells were starved of glucose for 4 h and supplemented with glucose for 30 min. a, c, d, e, f, g, i Each sample was subjected to immunoprecipitation with sWGA-conjugated agarose beads, anti-LARS1 antibody-conjugated beads or anti-myc 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.