Fig. 8. Chemotaxis of neutrophils to 9β1 integrin ligands. (A) Chemotaxis was assessed in a Boyden chamber with fluoroblock membranes (3.0 µm). Neutrophils isolated from human blood were labeled with calcein by incubation at 37°C for 30 minutes. Neutrophils were added to the upper chamber, and integrin ligands such as Y9A2 (5 µg/ml), VLO5 (1 µM) and mNGF (5 µg/ml), or 2% FBS were applied to the lower chamber. The plate was incubated at 37°C for 2 hours. Migration level was estimated as described in Fig. 5. Error bars indicate s.d. from three separate experiments. All chemoattracted groups were significantly different in comparison with random migration (P<0.001). (B) Inhibition of chemotaxis of neutrophils to mNGF. Migration was assessed after 4 hours. Random migration is represented by bar a and control migration to mNGF without inhibitors by bar b. Inhibition of chemotaxis of neutrophils to mNGF (5 µg/ml) by control mouse IgG (10 µg/ml), c; Y9A2 (10 µg/ml), d; disintegrins eristostatin (1 µM), e and VLO5 (1 µM), f. Error bars indicate s.d. from three separate experiments. ^*Significant difference (P<0.001) in relation to the group treated with 5 µg/ml of mNGF alone (b).