The Transwell invasion assay was performed as described previously [12 (link)]. Briefly, a total of 4 × 104 cells suspended in 100 μl of serum-free medium were added to the upper chambers of the Transwell system (24 wells, 8-mm pore size; Corning Costar, Lowell, MA, USA) coated with 2 mg/ml Matrigel (BD Biosciences). RPMI-1640 containing 20 % FBS and baicalein (20 and 40 μM) was then added to the lower chamber. After 24 h, the non-invaded cells in the upper chamber were gently removed with a cotton swab, and the cells attached to the lower surface were fixed with precooled methanol and stained with 0.1 % crystal violet. Five fields of each chamber were randomly selected, and the cell numbers were counted under a microscope.
For the migration assay, the cells were seeded into upper chambers that were not coated with Matrigel. The following steps in the assay were the same in the invasion assay. After 24 h, the cells on five randomly selected fields in the lower surface were counted, and the cell numbers were then subjected to statistical analysis.
For the migration assay, the cells were seeded into upper chambers that were not coated with Matrigel. The following steps in the assay were the same in the invasion assay. After 24 h, the cells on five randomly selected fields in the lower surface were counted, and the cell numbers were then subjected to statistical analysis.
Free full text: Click here
