Cells were treated with vehicle or various concentrations of CYD19 for 48 hours, and equal numbers (2 × 105 cells per well) of the cells were seeded in FBS-free DMEM culture medium in the presence of vehicle or various concentrations of CYD19 in the upper chambers of transwell inserts with an 8-μm pore size (BD Biosciences, #354480). The lower chambers were filled with 1 ml of complete medium supplemented with 10% FBS. Cells were allowed to invade the bottom chamber for 12 or 18 hours. Noninvading cells in the upper surface were removed, and invaded cells on the lower surface were fixed with 90% methanol and stained with 0.1% crystal violet for 5 min. The stained cells were photographed and quantified.

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