Figure 2.

Notch1 is involved in prolonged CdCl₂-induced EMT, stress fiber formation, and high cell motility in A549 cells. A, cells were incubated with 0, 1, 5, 10, or 20 μm CdCl₂ (Cd) for 10 weeks. Cell lysates were subjected to Western blotting using the indicated antibodies. B, prolonged 20 μm CdCl₂-exposed A549 cells (Cd) were transfected with control siRNA, Notch1 siRNA-1, or Notch1 siRNA-2. Cell lysates were subjected to Western blotting using the indicated antibodies. C, prolonged 20 μm CdCl₂-exposed A549 cells (Cd) were treated with 0.1% DMSO or 50 μm DAPT for 16 h. Cell lysates were subjected to Western blotting using the indicated antibodies. D, prolonged 20 μm CdCl₂-exposed A549 cells (Cd) were transfected with control siRNA, Notch1 siRNA-1, Notch1 siRNA-2, Snail siRNA-1, or Snail siRNA-2. Cell lysates were subjected to Western blotting using the indicated antibodies. Immunoblots shown are representative of at least three independent experiments. e and f, prolonged 20 μm CdCl₂-exposed A549 cells (Cd) were transfected with control siRNA, Notch1 siRNA-1, or Notch1 siRNA-2. Transfected cells were stained with phalloidin-FITC for F-actin and DAPI for DNA (E) and used for wound healing assays (F). Scale bar, 20 μm (e), 250 μm (F). Results of quantification of the area covered by cells are also shown (F). Each value (mean ± S.D. (error bars) of three experiments) was expressed as a percentage of the area covered. **, p < 0.01, significant difference between the samples.