Ecm0542d

hTERT-immortalized human mammary epithelial cells (IMEC) and XD cells were cultured at 37 °C and 5% CO₂ in 1:1 DMEM/F-12 medium (gibco #11320-074) supplemented with insulin (Clonetics, MEGM SingleQuots #CC-4136), EGF (Clonetics, MEGM SingleQuots #CC-4136), bovine pituitary extract (Clonetics, MEGM SingleQuots #CC-4136), hydrocortisone (Clonetics, MEGM SingleQuots #CC-4136) and 100 ng/ml cholera toxin (Sigma #8052). IMEC-MYC, IMEC-PIK3CA^H1047R, IMEC-P53DD, and IMEC-RAS were generated by transducing IMEC with pMXs-c-Myc, PGK-PIK3CA^H1047R, pBABE-puro-RAS V12, and MSCV-p53DD-iGFP vector, respectively. IMEC-MYC-7TGP cells were generated by transduction of IMEC-MYC with 7TGP vector. T47D and MCF7 cells were cultured at 37 °C and 5% CO₂ in DMEM high glucose (Euroclone #ECB7501L) supplemented with 10% fetal bovine serum (Euroclone #ECS0180L), 1 mM sodium pyruvate (Euroclone #ECM0542D) and 2 mM glutamine (Euroclone #ECB3000D). ZR751 cells were cultured at 37 °C and 5% CO₂ in RMPI Medium 1640 (gibco #31870-025) supplemented with 10% fetal bovine serum, 2 mM glutamine and 1 mM sodium pyruvate. MCF7-MYC, T47D-MYC, and ZR751-MYC were generated by transduction with pMXs-c-Myc.

LNCaP, DU145, PC3, and 22Rv1 PCa cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were cultured in a humidified incubator in 5% CO2 at 37 °C. DU145 and PC3 cells were cultured in RPMI-1640 medium (#30-2001, ATCC) supplemented with 10% Fetal Bovine Serum (FBS), (certified, One Shot™ format, Thermo Fisher Scientific, Waltham, MA, USA); 22Rv1 cells were cultured in Dulbecco’s modified Eagle’s Medium (DMEM) with 10% FBS; LNCaP cells were cultured in RPMI 1640 with 10% FBS supplemented with 10 mM HEPES (#ECM0180D, Euroclone, Mlan, Italy) and 1 mM Sodium Pyruvate (#ECM0542D, Euroclone, Mlan, Italy). All media were supplemented with antibiotics (150 U/mL penicillin, 200 U/mL streptomycin) (#ECB3001D, Euroclone, Mlan, Italy) and 2 mM Glutamine (#ECB3000D, Euroclone, Mlan, Italy). All cell lines were routinely tested using a PCR Mycoplasma Detection Set (#6601, Takara). Cell line authentication (STR) was carried out.