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6 protocols using Bortezomib

Piritrexim isethionate was a generous gift from Dr. Neil Clendeninn, former Burroughs Wellcome, Research Triangle Park, NC.; Other antifolates were obtained as previously described.3 (link) Doxorubicin, tunicamycin, cytosine β-D-arabinofuranoside (Cytarabine) and VP16 (etoposide) were from Sigma-Aldrich (St. Louis, MO, USA); Bortezomib (Velcade) was from AdooQ Bioscience (Irvine, CA, USA).
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Heregulin β-1 (Sigma, H7660) was used at a working concentration of 20 ng.mL−1. For lysosome and proteasome inhibition, cells were treated for 4 h with 50 nM of bafilomycin A1 (Calbiochem, 196000) and 1 µM of bortezomib (Adooq Bioscience, A10160), respectively. CHX (Sigma, 01810) was used for translation inhibition at 25 µg.mL−1. For targeted inhibition of intracellular signaling proteins, trametinib (Adooq Bioscience, JTP-74057, [100] nM), rapamycin (Santa Cruz Biotechnology, sc-3504A, [100] nM), buparlisib (Adooq Bioscience, AT11016, [500 nM]), selumetinib (Adooq Bioscience, A10257, [1 µM]), SCH772984 (Adooq Bioscience, A12824, [1] µM) and MK-2206 (Adooq Bioscience, A10003, [2] µM) were used for the indicated time points.
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KIRA8 was house-made as previously reported [10 (link)]. Bortezomib (CAS#179324-69-7) was purchased from AdooQ BioScience (Irvine, CA, USA), thapsigargin (CAS#67526-95-8) from Sigma-Aldrich (St. Louis, MO, USA), and nilotinib (CAS#641571-10-0) from MedChemExpress (Monmouth, NJ, USA). Two PERK inhibitors, GSK2606414 (CAS#1337531-89-1) and AMG PERK 44 (AMG; CAS# 1883548-84-2), and dominant Polo-like kinase 2 (PLK2) inhibitor TC-S 7005 (CAS#1082739-92-1) were purchased from TOCRIS Bioscience (Bristol, UK). All reagents were dissolved in dimethyl sulfoxide (DMSO) to a final concentration of 10 mM as a stock solution and stored at −30 °C.
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Doxorubicin was purchased from Sigma-Aldrich (Sigma, UK), bortezomib from AdooQ Bioscience (York, UK), bromodomain inhibitor JQ1 from APExBio (York, UK), and ABT199 from Active Biochemicals (Hong Kong, China).
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The culture medium RPMI 1640 and DMEM, Penicillin/streptomycin (pen/strep) (10,000 U/mL; 10 mg/mL) and trypsin-EDTA (0.05% Trypsin, 0.02% EDTA in Phosphate Buffer Saline) were related from PAN-Biotech (Aidenbach, Germany). Bortezomib and TMP269 were ordered from AdooQ (AdooQ Bioscience, Irvine, California, USA). Both compounds were dissolved in DMSO at 10 mM. Further dilutions were made with the appropriate culture medium. A maximum concentration of 1% DMSO was used in all experiments.
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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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