Figure 2. Epitope class 1 and 2 antibodies inhibit c-Met activity in S114 cells and colony formation in soft agar. (A) S114 cells were treated with 10 µg/ml of CE-310393 (class 1), CE-310083 (class 2), or AF276 (R&D Systems) for between 15 min and 24 h. Cells were lysed and western blots were performed using an antibody to Y1234/1235 in the kinase domain to measure pY-Met levels and sc-10 to assess total Met protein. (B) Inhibition of S114 growth in soft agar. S114 cells were plated in soft agar with varying concentrations of CE-310393, CE-310083 or 10 μg/ml control anti-KLH antibodies and grown for 10 d before colony number was determined. Values represent means ± SEM. Results of Student’s t-test are indicated: *p < 0.05, **p < 0.01, ***p < 0.001