Figure 6

Effect of TPL and ATF on cell migration. (A) Transwell assay. HUVECs and HCT116 cells were treated with TPL (10 ng/mL) and/or ATF (10 nM). After 16 h pretreatment and 9 h incubation in the upper chamber, the cells migrating to the lower membrane were stained and counted in five fields with a magnification of × 100. N = 3, bar = 50 μm. The experiments were carried out in triplicate and representative data are shown. (B) Effects of TPL and ATF on uPAR, FAK and p-FAK protein expression. HCT116 cells were treated with TPL (10 ng/mL) and/or ATF (10 nM). Twenty-four hours later, cells were harvested for Western blotting analysis using indicated antibodies. The level of GAPDH served as the loading control. Band intensities were calculated using software Image J. Relative intensities are also shown. Data represent mean values of triplicate samples. (C) HCT116 cells were treated with TPL (10 ng/mL) and/or ATF (10 nM). Twenty-four hours later, cells were harvested for RNA extraction and quantitative real time PCR using primers specific for human MMP-2, MMP-9, and GAPDH (internal control). Data represent mean values of triplicate samples. *p < 0.05, *p < 0.01.