FIGURE 5. LIBS exposure and ligand binding of disulfide-bonded and glycan wedge mutants.

A, LIBS exposure. 293T cell transfectants were stained with anti-LIBS antibodies, D3 or LIBS1, in the presence of 1 mM Ca²⁺/1 mM Mg²⁺ or 0.2 mM Ca²⁺/0.2 mM Mn²⁺, with or without 25 μM cyclo-RGDfV or 1 mg/ml Fn7–10. LIBS epitope expression is expressed as MFI of the D3 or LIBS1 staining as a percentage of MFI of AP3 mAb staining. B, Fn binding. C, fibronectin fragment Fn7–10 binding. D, fibronectin fragment Fn9–10 binding. In B–D, 293T cell transfectants were stained with 50 μg/ml fluorescently labeled ligands in the presence of 1 mM Ca²⁺/1 mM Mg²⁺ or 1 mM Ca²⁺/1 mM Mn²⁺ plus 10 μg/ml activating mAb, LIBS1, and subjected to flow cytometry. Binding is expressed as MFI of ligand staining as a percentage of MFI of Cy3-AP3 antibody staining.