Figure 9.

Effects of γ2pf and poly-l-lysine on cell adhesion, cell migration, and phosphorylation of integrin β4. (A and B) Effect on adhesion of EJ-1 (A) and A431 (B) cells to Pr-LN5. Plates (96-well) were precoated with 0.25 μg/ml (A) or 0.125 μg/ml (B) Pr-LN5 and then coated with the indicated concentrations of γ2pf (•) and poly-l-lysine (▴), followed by blocking with BSA. The adhesion of EJ-1 (A) and A431 (B) cells onto the plates was measured as described in Figure 2A. (C) Effect on migration of A431 cells. A431 cells were inoculated into each well precoated with 0.3 μg/ml Pr-LN5, and 0.4 μg of γ2sa or poly-l-lysine (PL) was added to the culture medium containing 1% FCS. After preincubation for 1 h, the cell migration speed was measured as described in Figure 3A. (D) Effect on integrin β4 phosphorylation of A431 cells. A431 cells were treated with 0.8 μg/ml γ2pf or PL and with 0.01 μg/ml EGF on the Pr-LN5 substrate. Integrin β4 present in the cell lysates was immunoprecipitated with the anti-integrin β4 mAb 3E1. The immunoprecipitates were analyzed by immunoblotting with the mAb against PY-20. Left, immunoblotting patterns. Right, quantitative analysis of the integrin β4 phodphorylation. Other experimental conditions are described in Figure 4 and Materials and Methods.