Transwell 24 well plates with 8μm diameter filters

Transwell migration and invasion assays were performed using transwell 24-well plates with 8μm diameter filters (Corning, NY, USA). For migration assays, cells were placed into the upper chamber with the noncoated membrane. For invasion assays, filters were precoated with 40μl of diluted ECM gel for 4h. Cells (1×10⁵) in 200μl of serum-free medium containing different concentrations of Rg3 (0, 25, 50, and 100 μg/ml) were placed in the upper chamber and 500μl 10% newborn calf serum was placed in the lower chamber. After incubated for 20h, cells were fixed in methanol for 15 min and stained with 0.1% crystal violet for 15 min. Then cells on the upside of the filters were removed with a cotton swab, and cells on the underside were examined and counted under a microscope at ×200 magnification.

Transwell 24-well plates with 8-μm diameter filters (Corning) were used to detect the migration and invasion abilities of cells. Approximately 1×10⁵ cells in 200 μl of a serum-free medium were placed in the upper chamber and 500 μl 10% fetal serum was placed in the lower chamber. After incubation for 20-24h, the cells on the underside of the filters were examined and counted under a microscope at 200X magnification.