Fig. 8.

Removal of single monosaccharide GalNAc-O-Ser/Thr causes the loss of UN1 detection. A, Scheme of the glycosidases used to remove specific glycan branches of the UN1 antigen. B, Total extract (100 μg) of HPB-ALL cells was left untreated (lane 1), digested with neuraminidase alone (lane 2), or neuraminidase followed by sequential digestion with β 1,4-galactosidase (lane 3), β-N-acetylglucosaminidase (lane 4), and β1,3-galactosidase (lane 5) or endo-α-N-acetylgalactosaminidase (lane 6). Digest aliquots (50 μg) were separated on 8% SDS-PAGE and analyzed by Western blotting with UN1, CD43 (C-20), or anti-γ-Tubulin antibodies. One representative experiment out of four is shown.