2.6. Transwell Migration

RWPE-2 prostate cancer and U87 glioma cells stably expressing miR-1227 were seeded on to the upper well of 8 µm transwell inserts, followed by the addition of 10% FBS containing media to the lower chamber. After 6 h of incubation, at 37 °C, the transwells were fixed with 4% paraformaldehyde and stained with crystal violet. A cotton swab was used to remove cells on the top of the filter. The stained filters were washed three times in deionized water, and the filters were imaged on an all-in-one Keyence microscope. Image J was used to quantify the intensity of the stained cells that migrated to the bottom of the transwell. Intensity values were normalized to the corresponding vector control.