Fig. 3

Effects of NCTD on the expression and activation of EGFR and c-Met on HCT116 (a) and HT29 (b) human colon cancer cells. The cells were seeded in 10-cm dishes for 24 h, and then cells were treated with NCTD or gefitinib (c) at 6.25, 12.5, 25, 50, and 100 μM, respectively. After 72 h of incubation, cells were harvested for western blot analysis. Cells were treated with 100 μM of NCTD for the indicated periods after which the cells were collected and prepared for western blot analysis (d). HCT116 cells were treated with 50 μM NCTD or 50 μM NCTD plus 0.2 μM MG132 for 48 h (e). The expression of EGFR or β-Actin protein was detected by western blot analysis. The numbers underneath the blots represent band intensities (normalized to the loading controls, means of three independent experiments) measured by the Image J software. The standard deviations were all within ±15% of the means (data not shown). β-Actin was used as equal loading controls. The experiments were repeated for three times