Cell Migration and Invasion Assays

To explore whether the expression of AEG-1 affected cell migration and invasion, Transwell migration and invasion assays were performed [40 (link), 47 (link)]. For the migration assay, exponentially growing cells (5 × 10⁵ cells/mL) were seeded into the upper chamber (BD Bioscience) in culture medium with 1% FBS, whereas for the invasion assay, the upper chamber was pre-coated with Matrigel (BD Bioscience) prior to adding the cells (5 × 10⁵ cells/mL), and then they were all incubated for 24 h at 37°C. The lower chamber was added with 600 μL RPMI 1640 containing 10% FBS. Then, the cells on the upper surface of the filters would be scraped off with swabs. After fixed the membrane with 4% paraformaldehyde and stained with crystal violet, the cells were taken photos under microscope. At least 5 microscopic fields were counted for each transwell filter.