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2 protocols using vorinostat

1

Targeted Drug Screening Protocol

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Bosutinib, masitinib and midostaurin (PKC412) were purchased from LC Laboratories (Woburn, MA, USA), ponatinib from Selleck Chemicals (Houston, TX, USA), piceatannol and pimozide from Sigma Aldrich (San Louis, MO, USA), and dasatinib, sorafenib, sunitinib, tozasertib, vorinostat, everolimus (RAD001), erlotinib, gefitinib, and lapatinib from ChemieTek (Indianapolis, IN, USA). Imatinib, nilotinib, and BEZ235 were kindly provided by Dr.E.Buchdunger and Dr.P.W.Manley (Novartis, Basel, Switzerland). Stock solutions of drugs were prepared by dissolving in DMSO (Merck, Darmstadt, Germany). A specification of targeted drugs is shown in Supplementary Table S1. RPMI 1640 medium and fetal calf serum (FCS) were from PAA Laboratories (Pasching, Austria), 3H-thymidine from Amersham (Buckinghamshire, United Kingdom), and the Annexin V-FITC Kit from eBiosciences (San Diego, CA, USA). Recombinant human (rh) stroma cell-derived factor-1 alpha (SDF-1α) and rh eotaxin were purchased from R&D Systems (Minneapolis, MN, USA), and rh interleukin-5 (IL-5) from BD Bioscience (San José, CA, USA). A specification of monoclonal antibodies (mAb) used in this study is shown in Supplementary Table S2.
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2

Identifying Anti-Neoplastic Drug Candidates

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A number of anti-neoplastic drugs were tested for their ability to inhibit growth of MM cells: the tyrosine kinase inhibitors (TKI) bosutinib, dasatinib, imatinib, sorafenib, sunitinib, and nilotinib, the ErbB-receptor inhibitors lapatinib, erlotinib, and gefitinib, the Aurora-kinase inhibitor VX-680, the HSP90 inhibitor 17AAG, the PLK-1 inhibitor BI2536, the pan-BCL-2 antagonist obatoclax, and the HDAC-inhibitor vorinostat were purchased from Chemietek (Indianapolis, IN, USA). The PI3 kinase/mTOR inhibitor BEZ235 was obtained from Selleck Chemicals (Houston, TX, USA). Stock solutions of drugs were prepared by dissolving in dimethylsulfoxide, DMSO (Merck, Darmstadt, Germany). RPMI 1640 medium and fetal calf serum (FCS) were purchased from PAA Laboratories (Pasching, Austria), and 3H-thymidine from PerkinElmer (Waltham, MA, USA). FITC-labeled CD34 monoclonal antibody (mAb) 581, PE-labeled CD34 mAb 581, FITC-labeled CD138 mAb MI15, PE-labeled CD138 mAb DL-101, PerCP-labeled CD45 mAb 2D1, APC-labeled CD38 mAb HIT2, PE-labeled and Alexa Fluor® 647-labeled active caspase-3 mAb C92-605 were purchased from BD Biosciences (San Jose, CA, USA). The PerCP-labeled CD20 mAb 2H7 and the APC-labeled CD27 mAb O323 were obtained from Biolegend (San Diego, CA, USA), and an Annexin V/FITC kit from eBioscience (San Diego, CA, USA).
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