EXPERIMENTAL MODEL AND SUBJECT DETAILS

Normal diploid IMR90 human fibroblasts were cultured according to the ATCC in low oxygen (2% O₂) in DMEM (4.5 g/L glucose, Corning cat# 10-017-CV) with 10% FBS supplemented with L-glutamine, non-essential amino acids, sodium pyruvate, and sodium bicarbonate. Experiments were performed on IMR90 between population doubling #25–35. Melanoma (SKMel28), pancreatic (PATU8902), colorectal (HT-29, SW620, and SW480) tumor cells and lentiviral and retroviral packaging cells (293FT and Phoenix, respectively) were cultured in DMEM (Corning, cat# 10–013-CV) with 10% FBS. ES2 ovary tumor cell line was cultured in RPMI 1640 with 10% FBS. TCCSUP bladder cancer cell line was cultured in MEM/EBSS glutamine supplemented with 10% FBS. All cell lines were cultured in MycoZap and were routinely tested for mycoplasma as described in Uphoff and Drexler (2005). All tumor cell lines included in this work express wild-type CDKN2A according to TCGA (Cerami et al., 2012; Gao et al., 2013). According to ATCC, TCCSUP harbors a mutation in RB1. All cell lines were authenticated using STR Profiling (Genetica DNA Laboratories).

Two-month old male SCID mice were purchased from Charles River Laboratories. All mice were maintained in a HEPA-filtered ventilated rack system at the Penn State College of Medicine animal facility. Mice were housed up to 5 mice per cage and in a 12-hour light/dark cycle. All experiments with animals were performed in accordance with institutional guidelines approved by the Institutional Animal Care and Use Committee (IACUC) at the Penn State College of Medicine.