FIGURE 6.

Rac1 inhibitor blocks Snail-induced scattering and motility, whereas ROCK1/2 inhibitor promotes scattering and motility of Slug cells. AsPC1-vector, AsPC1-Snail, and AsPC1-Slug cells were induced with doxycycline (2 μg/ml) for 24 h. A and B, the cells were then suspended in three-dimensional collagen gel (2 mg/ml), and fresh serum-containing medium supplemented with doxycycline and ROCK1/2 inhibitor Y27632 (10 μm) or Rac1 inhibitor NSC23766 (10 μm) was added every 2 days for 4 days. The effect on colony morphology was examined by phase contrast microscopy (A), and the percentage of scattered colonies per field was quantified (B). C and D, cells were also plated onto thin-layer type I collagen matrix overlaid with colloidal gold and allowed to migrate for 24 h, and the tracks were photographed (C) and quantified (D). Error bars in B and D indicate S.E. The results are representative of at least three independent experiments. *, p < 0.05. NS, not significant.