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. 2003 Sep 29;162(7):1281–1292. doi: 10.1083/jcb.200304018

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Copyright © 2003, The Rockefeller University Press

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Figure 3. — Ephrin-A1 inhibits HGF-induced MDCK cell scattering and directional cell migration. (A) Schematic illustration of SOIL assay. MDCK cells were cultured to reach near confluence on coverslips in 24-well plates and were transferred onto 6-well dishes precoated with either Fc or ephrin-A1–Fc. After 24–48 h of stimulation with 20 ng/ml HGF, cells were fixed and stained with crystal violet. After removing coverslips, the rings of cells that had scattered off the coverslips were photographed. (B) Immobilized ephrin-A1–Fc (but not Fc) inhibits HGF-induced MDCK cell scattering in a density-dependent manner. (C) High resolution image shows that coating with 2 μg/cm² ephrin-A1–Fc significantly inhibited HGF-induced cell scattering compared with Fc control. (D) Ephrin-A1 inhibits HGF-induced directional cell migration. 100 ng FN was dried on the underside of Transwell filter, and both sides of the Transwell were then coated with10 μg/ml collagen. MDCK cells were added to the upper chamber of the Transwell and allowed to migrate toward the lower chamber containing 20 ng/ml HGF and 1 μg/ml ephrin-A1–Fc or Fc overnight. Cells migrating through the filter were counted from six high power fields. Values represent means ± SD.

Figure 3. — Ephrin-A1 inhibits HGF-induced MDCK cell scattering and directional cell migration. (A) Schematic illustration of SOIL assay. MDCK cells were cultured to reach near confluence on coverslips in 24-well plates and were transferred onto 6-well dishes precoated with either Fc or ephrin-A1–Fc. After 24–48 h of stimulation with 20 ng/ml HGF, cells were fixed and stained with crystal violet. After removing coverslips, the rings of cells that had scattered off the coverslips were photographed. (B) Immobilized ephrin-A1–Fc (but not Fc) inhibits HGF-induced MDCK cell scattering in a density-dependent manner. (C) High resolution image shows that coating with 2 μg/cm² ephrin-A1–Fc significantly inhibited HGF-induced cell scattering compared with Fc control. (D) Ephrin-A1 inhibits HGF-induced directional cell migration. 100 ng FN was dried on the underside of Transwell filter, and both sides of the Transwell were then coated with10 μg/ml collagen. MDCK cells were added to the upper chamber of the Transwell and allowed to migrate toward the lower chamber containing 20 ng/ml HGF and 1 μg/ml ephrin-A1–Fc or Fc overnight. Cells migrating through the filter were counted from six high power fields. Values represent means ± SD.