*4.10. Migration and Invasion Assays*

Cell migration assay was performed using Falcon HTS FluoroBlok 24-well inserts (BD Biosciences, Franklin Lakes, NJ, USA) and the cell invasion assay was performed using the 24-well Collagen-based Cell Invasion Assay (Millipore, Beverly, MA, USA). Briefly, we added serum-free medium to rehydrate each insert, then replaced it with a serum-free suspension with equal numbers of cells in the upper chamber, followed by a 12- to 24-h incubation period to allow cells to migrate toward (i.e., invade) the lower chamber, which contained medium with 10% fetal bovine serum. After removal of the noninvading cells in the upper chamber, cells that invaded through the inserts were stained, lysed in extraction buffer, and transferred to 96-well plates for colorimetric readings at 560 nm.
