Aew541

All cell lines were cultured at 37°C in a humidified atmosphere containing 5% CO2. A673, SKNEP1, SKNMC, and the SKPNDW lines were grown in Dulbecco’s Modified Eagle’s Media (Life Technologies) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich). A673 cells were supplemented with 1 mmol/L of sodium pyruvate (Life Technologies). The CADOES1 and TC32 lines were cultured in RPMI1640 (Sigma-Aldrich) with 15% and 10% FBS, respectively. All cells were grown in the presence of 10 units/mL of penicillin, 10 μg/mL streptomycin and 30 μg/mL of L-Glutamine, PSQ (Thermo Fisher Scientific). Whole exome sequencing, RNA sequencing and STR genotyping of all Ewing sarcoma cell lines used in these studies and validated cell line identity.(20 (link)) Cell lines were regularly tested for Mycoplasma by PCR (Sigma Aldrich). All cell lines were passaged between 4–12 weeks between thawing and use in the described experiments. All cell lines were a kind gift from Todd Golub (Broad Institute, Cambridge, MA), Nathanael Gray (Dana-Farber Cancer Institute, Boston, MA). Ewing sarcoma cell lines were treated with the CDK4/6 inhibitor ribociclib and the IGF1R inhibitor AEW541 kindly provided by Novartis Oncology. Palbociclib (Cat. No S1116), linsitinib (Cat. No S1091), and GDC0941 (Cat. No S1065) were obtained from Selleck Chemicals. THZ1 was a kind gift from Nathanael Gray.