Pdgfr tyrosine kinase inhibitor 3

MDA-MB-435s-HM cells were infected with 20 MOI of pGC-FU-Luciferase lentiviruses to obtain a stable cell line and generate a lung metastasis mouse model as described above. Between 10 and 20 days after injection, 0.5 ml of each specific inhibitor or antibody [including TIMP-1 (ProSpec-Tany TechnoGene Ltd., Rehovot, Israel), PDGFR tyrosine kinase inhibitor III (Santa Cruz Biotechnology, Santa Cruz, CA, USA) or anti-human CD31 antibody WM59 (WM59 clone, eBioscience, Thermo Fisher Scientific)] to the most significant proteins identified by protein microarray analysis were intraperitoneally injected every other day at a concentration of 10 µg/ml, while the same volume of saline was injected into control mice. After treatment, six mice in each group were examined based on the bioluminescence of luciferase in the lung area every 4–6 days as described above. Moreover, the ratio of VEGFR1⁺CD133⁺ HPCs in the lung at different time points was examined following the protocol outlined above.

Human EPCs were incubated alone or with platelets in the presence or absence of 2 ug/ml FGF basic Antibody (R&D, Minneapolis, USA) or 12.5 ug/ml PDGFR Tyrosine Kinase Inhibitor III (Santa Cruz, Ca, USA. This concentration was determined by dose-response experiments using several concentrations). Growth factors inhibitors were added to platelets and/or EPCs for 30 minutes prior incubation.