β1 Integrin is involved in ephrin-A3-Fc inducing DC adhesion to fibronectin. Ninety-six-well plates were coated with 5 μg/mL fibronectin (left) or poly-l-lysine (PLL; right) overnight at 4°C. Nonspecific binding sites were blocked at rt for 1 hour using 1% BSA/PBS. Day-11 CD34+-derived DCs (1 × 105 cells in 100 μL serum-free medium) were plated in the presence of immunoglobulin (▵) or ephrin-A3-Fc (1-20μg/mL; □) and were centrifuged at 1200 rpm for 3 minutes. To test for the effect of the RGD peptide, cells were preincubated for 5 minutes with the peptide (10 μM; ○) before treatment with the fusion proteins. After washing and fixation, adherent cells were stained with crystal violet. Values represent mean ± SD A570 absorbances from triplicate wells. Results are representative of 3 independent experiments.
