Fig. 2

Analysis of serum glycoproteins and N-glycosylation intermediates in fibroblasts. a SDS PAGE and Western Blot of serum proteins from a control subject (N), a patient diagnosed with phosphomannomutase 2-deficiencey (PMM2-CDG) and the patient described here (P). Antibodies towards α1-antitrypsin (AAT), transferrin (TF), haptoglobulin (HAPTO) and orosomucoid (OROSO) were used to probe the blots. The numbers to the right of the images indicate glycoforms with 0, 1 and 2 N-glycans. b Fibroblasts derived from a control subject (N) and the patient (P) were metabolically radiolabeled with [2-³H]mannose and oligosaccharides derived from dolichol linked oligosaccharides (DLO) and glycoproteins (NLO) were resolved by TLC. The abbreviations used are: Man_4-8GlcNAc₂; M_4–8, Glc_0-3Man₉GlcNAc₂; G_0-3M₉. c Fibroblasts derived from three normal subjects (Ctrls) and five patients diagnosed with the indicated CDG I subtypes were metabolically radiolabeled and, the quantity of radioactivity associated with DLO and pronase-solubilised glycoproteins (N-glycan) was assayed by scintillation counting. The results are expressed as ratio of radioactivity associated with N-glycans/radioactivity associated with DLO