Fig. 1: UGDH correlates with poor prognosis and possesses high O-GlcNAcylation in the tumor.

A TCGA analysis shows that UGDH expression is significantly upregulated in multiple cancer tissues, especially in hepatocellular carcinoma, breast invasive carcinoma, and NSCLC. B UGDH expression in NSCLC is higher in patients with stage III-IV than stage I-II according to the Barcelona criteria. C Kaplan–Meier analysis demonstrates that high levels of UGDH expression are associated with shorter survival outcomes in NSCLC. D UGDH has an evolutionarily conserved amino acid sequence across different species, which comprises residues of potential O-GlcNAcylation. E, F Analysis of UGDH glycosylation in HEK293T cells overexpressing Flag-tagged UGDH under co-transfection of HA-OGT or TMG treatment. G Immunoblotting analysis of the glycosylation ratio of UGDH using enzymatic labeling coupled with alkyne PEG-5kD. H, I The co-immunoprecipitation of OGT/OGA with UGDH. Immunoblotting analysis of UGDH glycosylation after different nutrient stimuli, including glucose (J), H₂O₂ (K), glutamine (L) and EGF (M). N Immunoblotting analysis of UGDH expression and O-GlcNAcylation levels from primary NSCLC tissues and the matched adjacent tissues. Immunoblotting quantification of UGDH expression (O) and O-GlcNAcylation levels (P). Q Pearson’s correlation test was used to analyze the relationship between the levels of UGDH expression and UGDH O-GlcNAcylation. R Identification of O-GlcNAcylation sites on UGDH by high-resolution tandem mass spectrometry spectrum analysis. S Verifying the UGDH glycosylation sites using alanine mutations. Results are representative of three biological replicates.