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5 protocols using lenvatinib

1

Lenvatinib Quantification Protocol

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Lenvatinib was purchased from AdooQ BioScience LLC (Irvine, CA, USA). Internal standard propranolol (C16H21NO2) and acetonitrile (CH3CN, LC-MS/MS hypergrade) were purchased from Merck, Ltd. (Darmstadt, Germany). Methanol (CH3OH, LC-MS/MS hypergrade) and formic acid (HCOOH, LC-MS/MS hypergrade) were obtained from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). High-purity deionized water was obtained from an Elix 10-Milli-Q Plus water purification system (Millipore, Ltd., Eschborn, Germany).
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2

Hepatoma Cell Line Culture and Drug Treatments

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Human hepatoma cell lines HepG2 and Huh7 were used as previously described [9 (link),40 (link)]. Cells were maintained in Roswell Park Memorial Institute medium (RPMI1640) (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% fetal calf serum (FCS) and 200 U/mL penicillin and 200 μg/mL streptomycin at 37 °C in a 5% CO2 atmosphere. Sorafenib, regorafenib, and lenvatinib were purchased from AdooQ Bioscience (Irvine, CA, USA), Cayman Chemical (Ann Arbor, MI, USA), and ChemScene LLC (Monmouth Junction, NJ, USA), respectively.
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3

Inhibition of MEK and EGFR Pathways

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E7080 (lenvatinib) was purchased from AdooQ BioScience (Irvine, CA, USA). Selective MEK inhibitors, U0126 for the in vitro study and AZD6244 (selumetinib) for the in vivo study, were purchased from MedChemExpress (Monmouth Junction, NJ, USA) and Selleck Chemicals (Houston, TX, USA), respectively.
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4

Lenvatinib Cytotoxicity Assay Protocol

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Lenvatinib was purchased from AdooQ Bioscience LLC, and a solution of Lenvatinib was prepared by dilution with DMSO and stored at -20˚C. RPMI-1640 was obtained from Gibco (Thermo Fisher Scientific, Inc.). Trypan blue was purchased from MilliporeSigma. DMEM, minimum essential medium (MEM), and FBS were obtained from Wako Pure Chemical Industries, Ltd. Penicillin-streptomycin was obtained from Invitrogen (Thermo Fisher Scientific, Inc.). A cell cycle phase determination kit was obtained from Cayman Chemical Company, a protease inhibitor cocktail from iNtRON Biotechnology, and a Proteome Profiler Human Angiogenesis Antibody Array Kit from R&D Systems, Inc.
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5

Cell-based Assay of Multi-Targeted Kinase Inhibitors

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Regorafenib, M-2, M-5, and elacridar were purchased from Toronto Research Chemicals (North York, ON). Ko143 was from Sigma-Aldrich (St. Louis, MO). Imatinib mesylate was from Focus Biomolecules (Plymouth Meeting, PA). Afatinib, bortezomib, cabozantinib, carfilzomib, dabrafenib, dacomitinib, dasatinib, lapatinib, lenvatinib, pazopanib, ponatinib, ruxolitinib, tofacitinib, trametinib, vandetanib, and vemurafenib were from AdooQ Bioscience (Irvine, CA). Sorafenib was from LKT Laboratories Inc. (St. Paul, MN). Sunitinib was from Synkinase Pty Ltd. (San Diego, CA). Crizotinib was from LC Laboratories Inc. (Woburn, MA). Gefitinib and erlotinib were from Cayman Chemical Company (Ann Arbor, MI). Nilotinib was from ChemScene, LLC (Monmouth Junction, NJ). All other chemicals and reagents were of analytical grade and were obtained from commercial sources.
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