C met

Protein extraction and Western blot analysis were performed as previously described.18 (link) Representative images from 2 or 3 independent experiments are shown. The antibodies that were used included c-MET, E-cadherin, N-cadherin, Vimentin, ZEB1, Slug, Snail and GAPDH (Sigma Aldrich).
To detect c-MET expression in glioblastoma, IHC staining was performed on tumor tissue using methods described previously.17 (link) c-MET were scored by an IHC score based on staining intensity and percentage of positive cells within the whole tissue section.

Total protein extracted from tissues was subjected to electrophoresis using a 4–12% Bis-Tris gradient gel (Invitrogen, MA, United States) for 50 min. After transferring the gel to PVDF membranes (GE Healthcare, MA, United States), membranes were then blocked with Blocker Casein in PBS (Thermo Fisher, MA, United States) at room temperature (45 min), followed by the incubation with primary antibodies specific to c-Met (Sigma, MO, United States), phosphorylated c-Met (Cell Signaling Technology, MA, United States), c-Jun (Cell Signaling Technology, MA, United States), phosphorylated c-Jun (Cell Signaling Technology, MA, United States), and GAPDH (Cell Signaling Technology, MA, United States) 4 °C (overnight), respectively. Membranes were washed three times TBST (Invitrogen, MA, United States) and then subjected to incubation with a secondary antibody specific to anti-rabbit IgG (Cell Signaling Technology, MA, United States) at room temperature (1 h). Immuno-stained membranes were exposed to Super Signal West Femto Maximum Sensitivity Substrate (Thermo Fisher, MA, United States) or Super Signal West Pico Chemiluminescent Substrate (Thermo Fisher, MA, United States), and signal intensities were visualized using ImageQuant Las 4000 (GE Healthcare, MA, United States), followed by the quantification using ImageJ software (NIH).