4.5. Transwell assay

Transwell chamber (pore size 8 μm; Corning) was used to determine the invasion of LoVo cells according to previous report. 29 LoVo cells (1 × 10⁴) were placed in serum‐free DMEM (Gibco) in the upper chamber coated with matrigel. And the lower chamber was added with DMEM medium supplemented with 10% fetal bovine serum. Twenty‐four hours later, cells in the lower chamber were collected and stained with crystal violet. The number of invasion cells was counted using a microscope (Nikon).