β tubulin

The term “eNK1 monomer” refers to the engineered NK1 variant M2.2 D127N, and “eNK1 dimer” refers to the disulfide-linked dimer cd D127N from our previous work [29 (link)]. HGF was from R&D Systems. FGFb was from GIBCO. BaF3-MET cells were provided by Patrick Ma, Case Western Reserve University. BaF3-MET growth media was RPMI 1640 medium with Glutamax (Invitrogen) with 10% fetal bovine serum (FBS) (American Type Culture Collection), 1% penicillin-streptomycin, 0.5 ng/mL mouse interleukin-3 (R&D Systems), and 1 mg/mL Geneticin (GIBCO). Human umbilical vein endothelial cells (HUVECs) and the EGM-2 BulletKit were purchased from Lonza Walkersville Inc. A549 cells were grown in Dulbecco’s Modified Eagle Media (Invitrogen) with 10% FBS and 1% penicillin-streptomycin. The PE-conjugated anti-FLAG antibody for binding assays was from Prozyme. Primary antibodies for Western blot analysis were all from Cell Signaling except for β-tubulin (Covance) and c-MET (Santa Cruz Biotechnology). All secondary antibodies were purchased from Jackson ImmunoResearch, and chemoluminescent substrate was from ThermoScientific. NP-40 buffer was composed of 20 mM Tris pH 8.0, 137 mM NaCl, 10% glycerol, and 1% IGEPAL/NP40. All cells and reagents for the PathHunter Assay were from DiscoveRx Corporation.