Cgp 57380

The following 43 reagents were used in this study: AMG 102 and panitumumab (Amgen); cercosporamide (BioAustralis); AKT inhibitor V (Calbiochem); AS-252424, bisindolylmaleimide, CGP 57380 and imatinib (Cayman Chemical); CTX ILK inhibitor (CRC); Avastin (bevacizumab) (Genentech/Roche); 4EGI-1, ABT-737, ABT-737 enantiomer, pazopanib, and retinoic acid (Santa Cruz Biotechnology); erlotinib, lapatinib, sorafenib, and temsirolimus (Scientifix); bortezomib and CYT387 (Selleck Chemicals); AKT-I-1/2, axitinib, bicalutamide, cilostazol, cyclopamine, DAPT, dasatinib, docetaxel, doxorubicin, floxuridine, fluorouracil, goserelin, ifosfamide, PD98059, ribavirin, salinomycin, SU11274, sunitinib, Tamoxifen, and vinblastine (Sigma); NSC 7908 (Tocris); and temozolomide (Wyeth).

HEK293T cells were cultured in high-glucose Dulbecco’s Modified Eagle Medium (DMEM, Merck, Darmstadt, Germany), and CHO-K1 (CHO) cells in DMEM/F-12 (Merck). Both cell culture media were supplemented with 10% fetal bovine serum, 2 mM l-glutamine, and 1% penicillin/streptomycin (all Thermo Fisher Scientific). Transfection of generated CDH1 expression constructs was done using the Lipofectamine 3000 reagent (Thermo Fisher Scientific). For stable expression, transfected CHO cells were treated with 20 µg/ml puromycin (Thermo Fisher Scientific) for 21 days. GFP-positive cells were isolated using a MoFlo XDP cell sorter (Beckman-Coulter, Brea, MA, USA). To inhibit MAPK interacting serine/threonine kinase 1 (MNK1), HEK293T and CHO cells were treated with 75 or 200 µM CGP 57380 (Cayman Chemicals, Ann Arbor, MI, USA) for 24 or 72 h.

Compounds and growth factors used to treat cells were: Rapamycin, TGFβ, HGF (EMD Millipore, Billerica, MA), Activin (eBioscience, San Diego, CA), BMP4, Nodal, Torin1 (R&D Systems, Minneapolis, MN), Insulin, Transferrin, and Selenium (ITS) (Roche, San Francisco, CA), SU11274, AG490 (Sigma Aldrich, St. Louis, MO), and CGP57380 (Cayman Chemicals, Ann Arbor, MI). U0126 was obtained from Promega (Madison, WI). A TGFβ Type I Receptor kinase inhibitor (616451) was purchased from EMD Millipore (Billerica, MA).