6. Scratch wound healing assay and invasion assay

AGS cells and MKN28 cells transfected with siN-BLRs or siCT were re-seeded in 6-well culture plates. When cells reached an approximate 60% to 80% confluency, the bottom of each well was scratched using a P-20 tip. The width of the scratched cells was measured at 0 and 24 hours under a bright-field microscope. For the invasion assay, using the same cell line and conditions above, cells were replated on BD BioCoat transwells (BD Biosciences, San Jose, CA, USA) following the manufacturer’s protocol. After 24 hours, non-invading cells within the chamber were removed, and the upper layer of the trans-well was wiped with a cotton swab. The membrane of the bottom part of the upper chamber was fixed with 5% acetaldehyde buffer and stained with crystal violet solution. The invading cells on the membrane were counted under a bright-field microscope.