Evaluating Cell Migration Mechanisms

Wound-healing and transwell assays were used to evaluate migration. LX2 cells were inoculated into 6-well plate, and when they grew to 100% confluence, the cell monolayer was wrapped with a 200 μL pipette tip. The wound was photographed, which served as the 0-h time point. Cells were incubated in serum-free culture medium for 48 h, and wound healing was photographed. For the transwell assay, LX2 cells (2 × 10⁴ cells/200 µL) were seeded in the top chamber with 5% serum in a 24-well polycarbonate transwell filter (8 µm pore size, Corning Incorporated, USA), which was filled with 20% fetal bovine serum (700 µL) and placed in the lower chamber. After 48 h of incubation, cells grown in the polycarbonate transwell filter were fixed with 4% paraformaldehyde and stained with 0.1% crystal violet. The cells were wiped from the apical chamber with a cotton swab, and migrated cells were photographed with an inverted microscope [17 (link)]. The grouping and processing of cells are the same as CCK8 assay.