Figure 4

ADPGK finetunes glucose flux into O- and N-glycosylation. (a,b) Sugar precursors (UDP-GlcNAc, UDP-Gal, UDP-Glc, GDP-Man) of glycosylation. N = 4 independent experiments. (c, d) Representative immunoblots for GlcNAc-residues via monoclonal antibody against (c) O-GlcNAc and (d) the lectin VVL in lysates of control and ADPGK KO cells stimulated with PMA for different time periods. N = 3 independent experiments. (e) Abundance profile of changes in N-glycans in control and KO1 cells upon 24 h PMA/Iono stimulation and (f) scheme of changes in high mannose glycans, complex- and hybrid-type glycans in KO1 cells upon 24 h PMA/Iono activation analyzed via HILIC-FLR-MS. Mean of CTRs (TF-CTR and WT-CTR) and KOs (KO1, KO2, KO3) are shown. In all experiments stimulation was induced as described in the text using 10 ng/mL PMA and 10 µM Ionomycin. *p < 0.05. All images of blots represent cropped blots of appropriate protein size. For full length blots see Supplemental Fig. 3.