Hyperactive end joining repair mediates resistance to DNA damaging therapy in p53 deficient cells (Flow Data Part 1 of 3)

The purpose of this experiment was to assess whether TP53 deficient cells are more intrinsically resistant to radiation, thus giving them a survival advantage over cells that have p53.
Conclusion:
Our conclusions are that TP53 deficiency promotes increased survival in irradiated cells compared to WT RPE1 cells.
Notes:
Detailed methods: To assess this, we plated a mixed competition assay with p53 WT labelled with mCherry and p53 deficient RPE1 cells in a 1:1 ratio in a 96 well plate. We irradiated cells at different doses from 2-6 Gy, and assessed growth over time from Day 0 to Day 6. To perform this experiment in a high-throughput manner, we used the Intellicyte-iQ Platform to then monitor the mCherry + (WT cells) and mCherry - (TP53 deficient cells) over time. As controls we plated WT cells mixed with fluorescent mCherry positive WT cells, we also had non-irradiated controls for each of the harvest time points. In addition, positive and negative gating controls for mCherry gating was utilized with each experiment. Intellicyte-iQ was calibrated daily by the UNC Flow Cytometry Core Facility.