Cell adhesion assay

To determine the effect of MLE on cell adhesion, an assay using fibronectin-coated plate was performed as previously described (Matsuura et al., 2006[26]; Kwak and Ju, 2015[22]). Briefly, 1 μg/ml fibronectin (Sigma-Aldrich) and 0.5 % bovine serum albumin (Sigma-Aldrich) were sequentially added in 96-well plates with 2 h interval at 37°C. HCT116 and H1299 cells were mixed with serum complete media containing MLE (0-700 µg/ml) and seeded in the fibronectin-coated 96-well plates (1 × 10³ cells/well). After 2 h, unattached cells were washed out with PBS. Attached cells were incubated with 0.2 % crystal violet (in 20 % methanol) for 10 min at room temperature. After dissolving the stained cells by 1 % SDS (Sigma-Aldrich), adherent cells were quantified spectrophotometically using a plate reader (Bio-Rad Laboratories) at the wavelength of 540 nm.