Figure 1.

Prolonged CdCl₂ exposure induces EMT, stress fiber formation, and high cell motility in A549 cells. A, cells incubated with 0, 1, 5, 10, or 20 μm CdCl₂ for 14 weeks were seeded and allowed to proliferate for 5 days. Total cell numbers were counted each day using a cell counter. Each value is the ratio of the cell numbers at Day 0 and reflects the mean ± S.D. (error bars) of three experiments with duplicate assays in each experiment. B, cells were incubated with 0, 5, 10, or 20 μm CdCl₂ for 10 weeks, and their phase-contrast micrographs were taken. Scale bar, 100 μm. C, 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. Immunoblots shown are representative of at least three independent experiments. D, cells were incubated with 0 or 20 μm CdCl₂ for 10 weeks and stained with phalloidin-FITC for F-actin and DAPI for DNA. Scale bar, 20 μm. E, cells were incubated with 0, 10, or 20 μm CdCl₂ for 10 weeks and allowed to migrate for 20 h after scratching. Results of quantification of the area covered by cells are also shown (E). Each value (mean ± S.D. of three experiments) was expressed as a percentage of the area covered. Scale bar, 250 μm. **, p < 0.01, significant difference between the samples.