As mentioned above, the experiments include both a highly spatial setting where the virus was inoculated into an already established tumor, and a mixed setting where infected and uninfected cells were mixed before the tumor cells were placed into the mouse.
Shortly before implantation, the cells were placed into sterile phosphate-buffered saline (PBS).
Before starting measurements, cells were placed in a running DMEM medium (supplemented with 25 mM glucose, 2 mM glutamine, 1 mM sodium Pyruvate, and without serum) and pre-incubated for 1 h at 37 °C in atmospheric CO2.
Before hormonal treatments, cells were placed in DMEM-F12 medium (Invitrogen, Carlsbad, CA, USA), supplemented with 5% dextran- coated charcoal stripped fetal bovine serum (DCC FBS; Hyclone, Logan UT, USA), 2% antibiotics-antimycotics (Invitrogen), 1% sodium pyruvate (Invitrogen) and 10 μg/ml insulin (Sigma-Aldrich, St Louis, MO, USA).
Mononuclear cells were separated by Ficoll density gradient centrifugation (Ficoll-Paque™ PLUS, Cat. no. 17-1440-03; ABersham Biosciences AB, Uppsala, Sweden) and then washed and counted before 1.5 to 2.0 × 10 cells were placed on each glass slide.
Twenty-four to forty-eight hours before the experiment, the cells were placed in a 12-well plate, each well bearing a glass cover, at 60 70 % confluency.
Five days before these experiments, the cells were placed into complete medium without G418 supplement.
Five days before the experiments, the cells were placed into complete medium without puromycin supplement.
Before ChIP-kinetic analysis, cells were placed in DCC medium during three days, then serum deprived during 24 hr.
One hour before the beginning of experiment, cells were placed in a saline solution of composition: 130 mM NaCl, 3.1 mM KCl, 1.0 mM K2HPO4, 4.0 mM NaHCO3, 5.0 mM dextrose, 1.0 mM MgCl2, 2.0 mM CaCl2, 10 mM HEPES, 1.0 mM ascorbic acid, 0.5 mM myo-Inositol, 2 mM pyruvic acid, pH 7.3.
