Wound-healing and transwell assays

Cells were inoculated in 6-well plates for cell transfection. After the cell density reached 80-90%, the pipette head was used for scratching. The floating cells were washed with PBS buffer. Cultured cells were grown for 16 h to allow wound closure. The wound healing rates were measured and compared to the width at 0 h. Transwell assay was used to detect cell invasion ability. Cells were collected and suspended in serum-free medium. Six hundred microliters of medium containing 20% FBS was added to the 24-well plate, and about 200 uL cell suspension was added to the upper chamber with matrigel, and cultured for 16 h in an incubator at 37℃. The chamber was fixed in 4% paraformaldehyde and then stained in 0.1% crystal violet solution for 30 minutes, respectively. A cotton swab was used to wipe the cells and matrigel out of the chamber. The images were observed under a microscope, and the number of invasive cells was calculated using the Image J software.