Oci aml3

K562, OCI-AML3, THP-1, and HEK-293 cells were purchased from ATCC. HNT-34 cell line was purchased from DSMZ. Human STR profiling was confirmed in all cell lines. K562, OCI-AML3, THP-1, and HNT-34 cell lines were cultured according to supplier’s instructions in RPMI (ThermoFisher) supplemented with 10% FBS (Sigma) and 1% Penicillin/Streptomycin (ThermoFisher). HEK-293 cells were cultured in DMEM (ThermoFisher) supplemented with 10% FBS and 1% P/S.

THP-1 (Acute Monoblastic/Monocytic Leukemia), OCI-AML3 (Acute Myeloid Leukemia), Karpas299 (ALK-Positive Anaplastic Large Cell Lymphoma), MV4-11 (Acute Monoblastic/Monocytic Leukemia), K562 (Chronic Myelogenous Leukemia) and Lenti-X™293 T (human embryonic kidney cell) cell lines used in this study were purchased from American Type Culture Collection bank and ensured for mycoplasma-free before experiments. THP-1, OCI-AML3, Karpas299, MV4-11 and K562 were cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (Gibco, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco, USA). Lenti-X™293 T cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) (Gibco, USA) supplemented with 10% FBS. Cells lines were maintained in the incubator with 37 ℃ and 5% CO_2. MV4-11 and THP-1 cells stably expressing firefly luciferase (ffLuc) were established by transduction with the lentivirus simultaneously encoding firefly luciferase and puromycin resistance gene, and cultured in a complete medium added with 1ug/mL puromycin (Gibco, USA).

MV4-11, MOLM-13, OCI-AML2, and OCI-AML3 were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). MV4-11 and MOLM-13 expressed MLL fusion oncoprotein with FLT3-ITD. OCI-AML2 and OCI-AML3 expressed mutant NPM1c+ THP1 without FLT3-ITD or NPM1c + mutant, all cells were cultured in RPMI 1640 with 20% fetal bovine serum (Gibco).

Cell lines MV4-11, MOLM-13, HL-60, THP1, Kasumi-1, OCI-AML3 and OCI-AML5 were either purchased at the DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Heidelberg, Germany) or at the ATCC (American Type Culture Collection, Manassas, VA, USA) or authenticated by the Multiplex human Cell Authentication test (Multiplexion, Heidelberg, Germany). MV4-11, MOLM-13, HL-60 and THP1 cells were maintained in RPMI 1640 medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS Superior, Biochrom GmbH, Berlin, Germany). Kasumi-1 cells were cultured in RPMI 1640 medium supplemented with 20% FBS. OCI-AML3 cells were maintained in α-MEM medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 20% FBS. OCI-AML5 cells were cultured in α-MEM medium supplemented with 20% FBS and 10 ng/mL GM-CSF (PeproTech GmbH, Hamburg, Germany). All cells were maintained in a humidified incubator with 5% CO₂ at 37 °C. Primary AML cells were obtained after patient’s informed consent and approval of the study by the ethics committee (PV3469, Ethik-Kommission der Ärztekammer Hamburg). Cells were isolated from bone marrow using density gradient centrifugation and cultured as described elsewhere [43 (link)].

Four AML cell lines, OCI-AML3, HEL, KG-1 and HL-60, and two diffuse large B-cell lymphoma (DLBCL) cell lines, OCI-LY1 and OCI-LY19, were purchased from DSMZ (Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures). One DLBCL cell line, TOLEDO, was purchased from the American Type Culture Collection (ATCC). HEL, KG-1 and HL-60 cells were cultured in RPMI 1640 medium supplemented with 10% foetal bovine serum (FBS), l-glutamine and 1% penicillin–streptomycin (Gibco, Grand Island, NY, USA) at 37 °C and 5% CO₂. OCI-AML3 and OCI-LY19 cells were cultured in α-MEM supplemented with 20% FBS, l-glutamine and 1% penicillin–streptomycin (Gibco) at 37 °C in the presence of 5% CO₂. OCI-LY1 cells were cultured in Iscove Modified Dulbecco Media (IMDM) supplemented with 20% FBS, l-glutamine and 1% penicillin–streptomycin (Gibco, Grand Island, NY, USA) at 37 °C and 5% CO₂.

AML cell lines MOLM-16,NOMO-1, and OCL-AML3 were purchased from DSMZ-German Collection of Microorganisms and Cell Cultures (DSMZ no. ACC 555: MOLM-16, ACC 542: NOMO-1, ACC 582: OCI-AML3). MOLM-16 and NOMO-1 were cultured in RPMI-1640 medium (Gibco/Thermo Fisher Scientific, Waltham, MA, USA) and OCI-AML3 in α-MEM (with nucleosides; Gibco/Thermo Fisher Scientific) supplemented with GlutaMAX (Gibco CTS/Thermo Fisher Scientific), foetal bovine serum (20% for MOLM-16 and OCI-AML3; 10% for NOMO-1), and antibiotics.

Three human AML cell lines, Kasumi-1 (ATCC, Manassas, VA) with t(8;21) and heterozygous KIT N822K mutation, SKNO-1 (JCRB, Osaka, Japan) with t(8;21) and homozygous KIT N822K mutation, and OCI-AML3 (DSMZ, Brauschweig, Germany) with NPM1 mutation and DNMT3A mutation, were used in this study. Kasumi-1 cells were maintained in RPMI 1640 medium (ThermoFisher Scientific/GIBCO, Waltham, MA) containing 20% fetal bovine serum (FBS), SKNO-1 cells in RPMI 1640 medium with 10% FBS containing 10 ng/ml granulocyte-macrophage colony-stimulating factor (GM-CSF; R&D systems, Minneapolis, MN), and OCI-AML3 cells in αMEM medium (GIBCO) containing 20% FBS. Cultures were grown at 37 °C in humidified atmosphere containing 5% CO₂. These cell lines were authenticated (16-Markers STR) by the Food Industry Research and Development Institute, Hsinchu, Taiwan. Their genetic profiles were identical to reported information.
Cabozantinib-malate was purchased from Selleck Chemicals (Houston, TX) and dissolved in DMSO for storage at −20 °C. Cycloheximide (CHX), MG-132, Z-VAD-FMK, and puromycin were purchased from TargetMol (Boston, MA), Tocris Bioscience (Abingdon, UK), Abcam (Cambridge, MA), and GIBCO, respectively.