Cell migration was determined according to a previously reported method with modifications37 (link) with a 24-well invasion chamber containing an upper and lower chamber with an 8 μm pore in between (Corning Life Science, Acton, MA). Briefly, cells were harvested and resuspended in DMEM containing 1% (v/v) fetal bovine serum. Subsequently, 100 μl of cell suspension (1 × 105 cells in total) was added into the upper chamber while the lower chamber was filled with 600 μl of DMEM containing 10% (v/v) fetal bovine serum without or with 20 μM ATO. After 24 hours of incubation, adherent cells in the lower chamber were fixed with methanol and stained with 10% (w/v) Giemsa. The pictures of five randomly chosen areas were captured with a microscope at 100× magnification for quantification analysis.
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