Mono mac 6

AML cell lines MOLM-14, MV4–11 (with FLT3-ITD mutation), MonoMac 6 (with activating FLT3-V592A point mutation), and THP-1 (FLT3 wild type) were purchased from the American Type Culture Collection (ATCC). The short tandem repeat (STR) analysis was not done on leukemia cell lines. The primary leukemia cells were isolated from bone marrow or peripheral blood of AML patients under the auspices of the institutional (IRB approved) Tumor and Cell Procurement Bank at University of Maryland and Johns Hopkins University with informed patient consent in accordance with the Declaration of Helsinki. The mononuclear cells were isolated using Lymphocyte Separation Medium (Cellgro, Mediatech) according to manufacturer’s protocol. Primary cells were used either fresh or after viable freezing in FBS/5% DMSO. AML16, AML17, and AML18 cells were used after thawing. AML20 cells were used freshly collected without cryopreservation. All cells were cultured in RPMI 1640 medium (Life Technologies) supplemented with 10% FBS (HyClone, Thermo Scientific) and 200 mM L-Glutamine (Life Technologies).

THP-1, MV4–11, MonoMac-6, HL-60, and K562 cells were purchased from the American Type Culture Collection (ATCC, Manassas, VA). MOLM-14 cells were the kind gift of Dr. Mark Levis from Johns Hopkins University. All cell lines were grown at 37°C with 5% CO₂ atmosphere with Roswell Park Memorial Institute (RPMI) 1640 (Life Technologies, Carlsbad, CA) supplemented with heat-inactivated 10% (v/v) fetal bovine serum (FBS). Cell lines were grown and maintained following ATCC recommendations. We performed in-house karyotyping and molecular analyses on the cell lines and report them in Supplementary Table 2.
Primary human leukemia cells (primary AML) derived from patients were obtained through the institutional (IRB approved) Tumor and Cell procurement Bank at the University of Maryland. Primary AML cells, which had been previously isolated through ficoll separation and frozen viably, were thawed into room temperature RPMI 1640 with 10% FBS and supplemented with DNase I 20U/ml. Viable cell numbers were obtained using trypan blue exclusion, and cells were plated into 96-well dishes at 5×10⁴ cells/well in RPMI/10% FBS.