Fig. 3

Chemical labeling of O-GlcNAcylated proteins in TNBC cells. a Scheme for O-GlcNAc detection: chemoenzymatic labeling using GalT (N-acetylgalactosaminyl transferase mutant Y189L) and copper-catalyzed azide-alkyne cycloaddition (CuAAC) click chemistry. b–d Total cell O-GlcNAcylation measured with TAMRA-red fluorophore-alkyne click reaction. N = 2, representative images shown. Cells were maintained in High glucose = 4.5 g/L or Low Glucose = 1.0 g/L DMEM media for at least 3 days prior to analysis. e Band densitometry for total O-GlcNAc staining, presented as total band density of high/low glucose. f–h O-GlcNAc enrichment using biotin click labeling and streptavidin beads followed by TET1 immunoblot. N = 2, representative images shown. i Band densitometry for TET1 O-GlcNAc labeling, presented as total band density of high/low glucose