Wound-healing migration assay

CAL 27 cells were transfected by 20 nM L-siRNA-1, L-siRNA-2 or non-specific (NS) siRNA on Day 0. Cells were passed and transfected again on Day 2. (Note: NS siRNA-transfected cells were passed at 1:4, whereas L-siRNA-1- or L-siRNA-2-transfected cells were passed at 1:2 on Day 2.) Cells grew to 95–100% confluence on Day 4. The monolayer of cells were wounded with a 200-μL micropipette tip and washed thrice with PBS on 0 h (t = 0). Cells were then cultured in serum-free medium. Wound healing was photographed at 40 h (t = 40). The cell migration was expressed as distance migrated by cells.