Trans well migration chambers

Cell motility was measured using both wound-healing and transwell migration assays. Cells were grown to a confluent monolayer on 6-well plates, and wounds were made on the surface of the cultured cells using a 200-μm micropipette tip. After wounding, the detached cells were removed by washing with PBS. Phase-contrast images of the wound were captured 24 h after wounding using an inverted light microscope (Nikon, Tokyo, Japan). Wound areas were measured using ImageJ software (NIH).
Transwell migration assays were performed using transwell migration chambers (8-μm pore size; BD Falcon, Franklin Lakes, NJ, USA). The transfected cells were treated with trypsin/EDTA solution and resuspended as single-cell solutions in serum-free DMEM. Cells in serum-free DMEM were seeded into the upper chamber of each insert, and DMEM containing 10% FBS was added to the lower chambers. After incubation for 24 h, cells that migrated to the bottom surface of the insert were fixed in 100% methanol for 20 min, stained with Giemsa solution, and rinsed in PBS. Cells on the top surface of the insert were removed by wiping with a cotton swab. Images were captured using an inverted light microscope (Nikon), and the number of cells in each photo field was counted using ImageJ software (NIH).