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Suspensions were sub-cultured every 14 days by transferring 3 ml of cells into 30 ml of fresh medium.
Fifty mL of cells (2.0 × 107 cells/mL) were treated with formaldehyde to cross-link proteins to DNA.
Aliquots (1 mL) of cells grown in liquid medium were also sampled and stained with FDA to determine the percentage of live cells.
Resistance to ultraviolet irradiation was tested by placing 20 ml of cells in Petri dishes and exposing them to short wavelength UV light (254 nm) using the ultraviolet lamp for 120 s (Zhao et al. 2007).
After 24 hours, 15 ml of cells were harvested for RNA isolation.
On day 3, 0.5 ml of cells was diluted to 5 ml while maintaining OHT concentrations.
Cells were grown and harvested as described above, where 1 ml of cells were pooled and pelleted.
For treatments, 3 ml of cells at a density of 5×105 were seeded in 6-well microtiter plates.
After 8 hrs of induction, 1 mL of cells was collected and the OD600 was measured using a spectrophotometer (Thermo Scientific Biomate3).
Following 0.5, 1, 2 and 4 hours of induction at 30°C, 45 ml of cells were removed, pelleted and immediately subjected to the ChIP protocol.
When larger amounts of cells were required, light exposure was performed in a temperature-controlled glass container 2.2 cm optical path, with 50 ml of cells.
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