Cell migration assay

A549 and H226 cell migration was measured using a Transwell chamber with a polycarbonic membrane with 8-mm pore size in vitro. NSCLC cells were treated with different transfection for 24 h. Then, 1×10⁵ cells were seeded into the upper chamber and a complete culture medium with 20% FBS was added to the lower chamber as an attractant. After incubation for 24 h at 37°C, in a 5% CO₂ environment, the media were removed. The cotton swabs were used to remove the cells from the top chamber. Moreover, the 100% methanol was used to fix the cells migrated to the lower membrane and 0.1% crystal violet was used to stain the migrating cells. The number of cells was quantified by a microscope (Olympus, Tokyo, Japan).