Transwell migration/matrigel invasion assay

After transfection for 48h, A2780 cell migration and invasion were determined using Transwell Permeable Supports with a pore size of 8 μm (Cat.3428, Corning) without or with Matrigel (Cat.356234, BD Biosciences). After serum starvation for 24h, the transfected cells that were resuspended (5×10⁴ cells per well) in 200μL of serum-free media were plated in the upper chambers. The lower transwell chamber contained the chemoattractant (10% FBS). For the invasion assay, the upper transwell chamber was coated with Matrigel, and for the migration assay, the chamber was uncoated. After incubated at 37°C and 5% CO₂ for 24h, the upper surface of the membranes was then gently scraped using cotton swabs and washed with PBS to remove the stationary cells. Then, the membranes were fixed in 95% ethanol for 25min followed by staining with hematoxylin. The number of migrated cells was counted and averaged between ten random fields per well.