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2 protocols using daunorubicin hydrochloride

1

AML Cell Line Characterization Protocol

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All human AML cell lines were purchased directly from recognized repositories; Kasumi-1, MV-4-11, CMK, AML-193, M-07e, HL-60, ML-2, OCI-AML3, ME-1 and HEL from DSMZ (Braunschweig, Germany) and THP-1 from ATCC (Manassas, USA).
AZD6244, PD0325901, GSK1120212, CI-1040, TAK-733, AS703026, MEK162, cytarabine (cytosine arabinoside), daunorubicin hydrochloride (daunorubicin) and etoposide were obtained from Selleck chemicals (Houston, USA). ARRY-162 was purchased from ChemieTek (Indiana, USA). All compounds were dissolved in DMSO and stored at –20° C.
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2

Validating IDH Mutant Compound Screens

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Drug screening was done using IDH inhibitors AGI-5198 (Agios) or BAY-1436032 (Bayer) or the FDA-approved Oncology Drug Set II library (National Cancer Institute) containing 107 compounds. The compounds Gemcitabine hydrochloride (Sigma-Aldrich), Paclitaxel (Sigma-Aldrich), Teniposide (Santa Cruz Biotechnology, Inc), Daunorubicin hydrochloride (SelleckChem), Romidepsin (MedChemExpress), Dactinomycin (BioViotica), Regorafenib (MedChemExpress), Omacetaxine mepesuccinate (Sigma-Aldrich), and Marizomib (Sigma-Aldrich), were selected for the validation studies. Initial NIH compound screens were carried out in a 96-well format on 7 IDH mutant cultures and validation studies were performed in 384-well plates on all 12 IDH mutant cultures using the STARlet automated pipetting system (Hamilton). Serial dilutions of the selected compounds were added after 24 hours and viability was assessed by CellTiter GLO 2.0 after five days. For details see Supplementary Methods.
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