Figure 2
The MET receptor is activated by HGF, or APC- or Biotin-conjugated anti-MET antibodies. (A) Serum-starved U373-MG cells were treated either with HGF (50 ng/ml) for 7 minutes or left untreated (-HGF) as indicated. The cells were directly lysed in SDS buffer and proteins were detected by Western blotting with antibodies for the Y1234/Y1235-phosphorylated MET (p-MET, Y1234/5), total MET, phosphorylated AKT (p-AKT), total AKT, or actin (loading control), as indicated. (B) Serum-starved U373-MG cells were left untreated, or treated with APC-conjugated mouse IgG control antibody or APC-conjugated mouse monoclonal anti-MET antibody (each at 1 μg/ml) for 7 minutes. Cells were directly lysed in SDS buffer and proteins were detected by Western blotting with antibodies as in panel A. (C) The same as panel B except that the Biotin-conjugated goat control IgG or Biotin-conjugated goat anti-MET antibodies (each at 2 μg/ml) were used. (D) Serum-starved human glioblastoma cells T98G, LN229, and LN18 cells were treated with HGF or biotin-conjugated IgG or biotin-conjugated anti-MET antibodies as in panel A and C, or left untreated. The proteins were detected by Western blotting with antibodies as in panel A.
