Iscove modified dulbecco media (imdm)

AML cell lines HL-60 (ACC-3), and MonoMac-1 (ACC-252) were obtained from DSMZ. MDS-L^{34 (link),35 (link)} was kindly provided by Dr. Starczynowski (Cincinnati Children’s Hospital Medical Centre, OH, USA). AML cell lines were cultured in RPMI medium (Biowest) supplemented with fetal bovine serum (FBS, Lonza), 2 mM L-Glutamine (Lonza) and/or 0.1 mM non-essential amino acids (Lonza) according to manufacturers’ recommendations. Primary MDS and CMML blasts were cultured in IMDM (Biowest) supplemented with 3% heat-inactivated FBS, 2 mM L-Glutamine, 20% BIT 9500 Serum Substitute (StemCell Technologies), 5 ng/ml IL3 (Peprotech), 1 mM sodium pyruvate and 5 × 10⁻⁵ M β-mercaptoethanol (Sigma-Aldrich,) and 0.1 mM non-essential amino acids (Lonza).

Human A549 cells (RRID:CVCL_0023; Cat#ATCC CCL-185; established from lung carcinomatous tissue from a 58-year-old Caucasian male), MCF-7 (RRID:CVCL_0031; Cat#ATCC HTB-22; established from breast adenocarcinoma from a 69-year-old Caucasian female), and RKO (RRID:CVCL_0504; Cat#ATCC CRL-2577; colon carcinoma cell line) (all obtained from ATCC, USA), were grown in DMEM medium (Fisher Scientific; Cat#11625200) supplemented with 10% FBS (Fisher Scientific; Cat#11560636), 2 mM l-glutamine (Fisher Scientific; Cat#BE17-605E) and 100 units per mL of penicillin/streptomycin (Thermo Fisher; Cat#15140122) and incubated at 37 °C in 5% CO₂. HFF-1 cells (RRID:CVCL_3285; Cat#ATCC SCRC-1041) were cultured under the exact same conditions in IMDM (Biowest). Cells were routinely tested for mycoplasma by MycoAlert Mycoplasma Detection Kit (#Cat: 11650261, Thermo Fisher Scientific). No authentication was performed, since passage number 2 cells were thawed and used from the ATCC source in all the experiments. In LC50 determination, cells were seeded at a density of 4000 per well in 96-well plates and after a day of growth, treated with the molecules for 48 h. Thereafter cell viability was measured using CellTiter-Blue® Cell Viability Assay (Promega; Cat#G8081) according to the manufacturer protocol.

BxPC-3 and Capan-1 cell lines were purchased from the ATCC (Manassas, VA, USA). MIA PaCa-2, MCF7, SK-BR-3 and MDA-MB-231 cell lines were in possession of 4Cell Therapies S.A. and were authenticated by Eurofins Genomics (Ebersberg, Germany). Capan-1 cells were cultured in IMDM, 20% FBS and 1× pen/strep (Biowest) on collagen I (Merck, Darmstadt, Germany)-coated plates. The BxPC-3 cell line was cultured with RPMI 1640 with 10% FBS and 1 × pen/strep (Biowest). The SK-BR-3 cell line was cultured in McCoy’s 5a supplemented with 20% FBS and 1 × pen/strep (Biowest). All other cell lines were cultured in DMEM high glucose supplemented with 10% FBS and 1 × pen/strep (Biowest).