FIGURE 9.

Snail enhances cell migration toward FN, osteopontin, and the conditioned medium of osteoblasts. A, migration assay of MDCK/neo and MDCK/snail cells to various attractant. Cells (1 × 10⁶ cells/0.1 ml) were seeded into the upper wells. FN or osteopontin was added to the lower wells at a final concentration of 10 μg/ml. BSA was used at 0.1%. Conditioned medium of MC3T3 cells (CM MC3T3) or fresh medium containing 10% FCS was added to lower wells at 25% v/v. After 4 h of incubation at 37 °C, cells that had migrated to the lower surface of the filter were counted using the MTT assay. Data are the mean ± the S.E. for triplicate determinations. *, p < 0.05 versus MDCK/neo cells. B, inhibition of migration of MDCK/snail cell to osteopontin by function blocking integrin αVβ3 antibody. MDCK/snail cells were incubated with 25 μg/ml function blocking integrin αVβ3 antibody or control antibody in DMEM supplemented with 0.1% BSA for 20 min. Cells (8 × 10⁴ cells/0.1 ml) were then seeded into the upper wells. Five μg/ml osteopontin, 1 μg/ml FN, and 1% FCS in DMEM supplemented with 0.1% BSA were added to the lower wells. Migration assay was performed as described above. *, p < 0.05 versus control antibody-treated cells.