Cytokine and inhibitors; purified recombinant human HGF was obtained from Peprotech. The c-Met receptor inhibitor (PHA-665752 ≤ 1 μM), MAPK pathway inhibitor (PD98059 ≤ 40 μM), Arp2/3 complex inhibitor (CK-666 ≤ 200 μM) and CDC42 inhibitor (ML-141 ≤ 20 μM) were obtained from Cayman Chemical. The ROCK pathway inhibitor (Y27632 ≤ 7.5 μM) and FAK inhibitor (PF-562,271; 1 μM) were purchased from Merck. The PI3K pathway inhibitor (LY294002 ≤ 100 μM) and the Rac1 inhibitor (6-Thio-GTP≤50 μM) were obtained from Selleck Chemicals and Jena Bioscience, respectively. The functionally blocking β1-integrin antibody (≤10 μg/ml) and its mouse IgG1 isotype were purchased from Abcam.
Immunofluorescence; primary and secondary antibodies, rabbit anti-Met, and rat anti-α-tubulin were obtained from Invitrogen. Mouse anti-paxillin was obtained from BD Bioscience. Mouse anti–β1-integrin was obtained from Abcam. Alexafluor-conjugated secondary antibodies (488, 568, and 647 nm), Rhodamine phalloidin, DiO’, and DiOC18(3) (3,3′-Dioctadecyloxacarbocyanine Perchlorate) were all purchased from Invitrogen. The mitochondrial staining kit, red fluorescence-cytopainter, was purchased from Abcam. Vectashield mounting medium was obtained from Vector Laboratories Ltd., and DAPI was obtained from Invitrogen.
Immunofluorescence; primary and secondary antibodies, rabbit anti-Met, and rat anti-α-tubulin were obtained from Invitrogen. Mouse anti-paxillin was obtained from BD Bioscience. Mouse anti–β1-integrin was obtained from Abcam. Alexafluor-conjugated secondary antibodies (488, 568, and 647 nm), Rhodamine phalloidin, DiO’, and DiOC18(3) (3,3′-Dioctadecyloxacarbocyanine Perchlorate) were all purchased from Invitrogen. The mitochondrial staining kit, red fluorescence-cytopainter, was purchased from Abcam. Vectashield mounting medium was obtained from Vector Laboratories Ltd., and DAPI was obtained from Invitrogen.