Transwell membrane 0.8 μm pore size cell culture insert

Migration and invasion assays were performed using a transwell membrane (0.8 μm pore-size cell culture insert, FALCON, Abilene, TX, USA). A non-coated transwell membrane was used for the cell migration assay, and a transwell membrane coated with 2 mg/mL Matrigel was used for the cell invasion assay. A total of 2 × 10⁴ cells were used for the migration assays. For the invasion assays, 2 × 10⁵ cells in serum-free medium were seeded onto the upper chamber and 750 μL medium supplemented with 10% FBS was added to the lower chamber. After incubation for 16 h, cells that did not migrate through or invade the pores were removed with a cotton swab. Cells that exhibited migration and invasion (adhered to the lower surface) were fixed with 4% paraformaldehyde, stained with 0.1% crystal violet and counted using a light microscope in three randomly selected fields. The experiments were performed thrice in triplicate.

We performed migration and invasion assays using Transwell membrane (0.8 μm pore‐size cell culture insert, FALCON). For cell migration assay, we coated a non‐coated Transwell membrane and for cell invasion assay Transwell membrane with 2 mg/ml Matrigel (354234, Corning). For migration assays and invasion assay, we seeded the cells in serum‐free medium onto the upper chamber and added 750 μl growth medium with 10% FBS to the lower chamber. After incubation for 18 h, we removed the cells that did not migrate through or invade the pores with a cotton swab. We fixed the migration and invasion cells, which adhered to the lower surface with 4% paraformaldehyde and stained with 0.1% crystal violet. We counted the cells using a light microscope in three randomly selected fields. We performed the experiments thrice in triplicate.