FIGURE 2.

Monoclonal antibodies to regions in 120-kDa fragment block Ail binding. A, binding sites for the antibodies used in this study are indicated. Plasma Fn was adsorbed onto plastic wells (10 μg/ml). Antibodies directed against the N terminus of FN (B) and more central region (C) were added to Fn-coated wells 1 h prior to the addition of E. coli AAEC185 derivatives expressing empty vector and Ail. Bacteria were allowed to bind for an additional 1 h at 37 °C. Bound bacteria were stained with 0.01% crystal violet. The cells and crystal violet were solubilized and washed, and the cells were solubilized and read at A₅₉₅. E. coli expressing Ail without antibody treatment was set at 100% to normalize bacteria binding across multiple experiments (n = 6–9). *, p < 0.0005; **, p < 5 × 10⁻¹² relative to Ail-mediated binding with no mAb treatment (−). Error bars, S.D.