Figure 9
Biotin-conjugated anti-MET antibodies inhibited cell invasion. (A) An equal number of U373-MG cells were seeded to the top chamber of the Boyden chamber precoated with Matrigel. Cells were incubated in the complete medium and the bottom chamber also contained complete medium supplemented with biotin-conjugated control IgG or anti-MET antibodies (each at 2 μg/ml), or with APC-conjugated control IgG or anti-MET antibodies (each at 1 μg/ml). After 24 hours, cells that migrated to the bottom chamber (invaded through Matrigel) were stained, imaged by an EVOS light microscope, and quantified using the ImageJ software. Assays were performed in triplicates, and the means (with S.D.) are shown at the bottom. Statistical analysis was conducted (compared with the corresponding control in each subgroup) (***p < 0.001, **p < 0.01). (B) LN229 cells were assayed, similar to that described in panel A, except that the cells were allowed to invade for 32 hours. (C) U373-MG cells were starved for 9 hours and the equal number of cells were then seeded to the top chamber of the Boyden chamber with serum-starvation medium, and the bottom chamber contained the serum-starvation medium supplemented with HGF (50 ng/ml), PBS, anti-MET antibodies or control IgG as in panel A, or combination of HGF and antibodies as indicated. Invasion assays were carried out for 24 hours. The assay plates were then processed as in panel A and similarly quantified. Assays were performed in triplicates and the means (with S.D.) are shown on bottom. Statistical analysis was conducted (compared with the corresponding control in each subgroup) (***p < 0.001, **p < 0.01). (D) LN229 cells were assayed, similarly to that described in panel C, except that the cells were allowed to invade for 32 hours. Assays were performed in triplicates, and the means (with S.D.) are shown on bottom. Statistical analysis was conducted (compared with the corresponding control in each group) (***p < 0.001, **p < 0.01).
