Ons 76

Two different and well characterized cell lines were used in our study, DAOY cells that directly derived from primary MB and ONS-76, described as a more immature cell line with a primitive profile [54 (link)]. DAOY and ONS-76 human MB cell lines were purchased from the American Type Culture Collection, (Manassas, VA). DAOY cells were cultured in Eagle's minimal essential medium (EMEM) supplemented with 1% Penicillin/Streptomycin and 10% fetal bovine serum (FBS) (all from Sigma-Aldrich, Milan, Italy) while ONS-76 was cultured in RPMI 1640 (GIBCO Cell Culture, Burlington, Ontario) containing 10% FBS, 1% L-glutamine 2 mM and 1% Penicillin/Streptomycin (Sigma-Aldrich). Cells were treated with 4-HPR (a kind gift from Dr. James A. Crowell, Division of Cancer Prevention, National Cancer Institute, Bethesda, MD, and Dr. Gregg Bullard, McKessonBio, Rockville, MD) dissolved in absolute ethanol in a stock solution of 10 mM and used at indicated concentrations for different time periods.

The Daoy and ONS-76 cell lines were obtained from American Type Culture Collection (ATCC, Manassas, VA, United States). The Daoy and ONS-76 cells were maintained in RPMI 1640 medium supplemented with 10% FBS and 1% penicillin–streptomycin (HyClone, Logan, UT, United States). The cells were maintained at 37°C with 5% CO₂ under the condition of a humidified atmosphere.

Two human SHH-MB cell lines, Daoy and ONS76, were originally obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). Daoy cells were cultured in Eagle’s minimum essential medium (Cytiva, Marlborough, MA, USA) supplemented with 10% fetal bovine serum (FBS) (Cytiva) and 1% penicillin-streptomycin (PS) (Cytiva). ONS76 cells were cultured in RPMI 1640 medium supplemented with 10% FBS and 1% PS. HEK293FT cells obtained from Invitrogen (ThermoFisher Scientific, Waltham, MA, USA) were cultured in high-glucose Dulbecco’s modified Eagle medium (DMEM) (Cytiva) supplemented with 10% FBS, 1% PS, and 1% non-essential amino acids (Cytiva). Cells were incubated at 37 °C in 5% CO₂ and were routinely passaged upon 90% confluence.