FIGURE 3.
CTD110.6 recognizes both O-GlcNAc and terminal β-GlcNAc on cell surface glycoproteins. A, N-glycans of glycoproteins synthesized in CHO glycosylation mutants cultured in the presence (predicted (28, 29)), or absence (26, 27), of swainsonine. B, binding of L-PHA (upper) and mAb CTD110.6 (lower) to fixed cells. Lec2 and Lec8 cells were cultured with or without 5 μg/ml swainsonine (SW) for 4 days. Cells (4 × 105) were incubated with 20 μg/ml fluorescein-labeled L-PHA or 40 μg/ml CTD110.6 on ice for 20 min followed by 10 μg/ml Cy5-conjugated anti-mouse IgM antibody. Gray profiles, secondary Abs; thin line, control cells; bold line, swainsonine-treated cells. Representative results are from two independent experiments.