Glycosylase selectivity of ATA in MCF7 cells

To determine if ATA inhibits removal of uracil, hypoxanthine, 8oxodG opposite dC, dA opposite 8oxodG, and T opposite dG.
Conclusion:
ATA inhibits uracil removal more strongly than other glycosylase substrates. ATA does not inhibit T:G repair or repair of dA oposite 8oxodG. It does inhibit repair of hypoxanthine and 8oxodG opposite dC
Notes:
Cells were transfected with plasmid cocktails for measuring DNA repair. The "Undamaged cocktail" contained BFP, GFP, mOrange, and mPlum plasmids. "Damaged 1" cocktail contained uracil (BFP), hypoxanthine (GFP), 8oxoG opposite dC (mOrange) and mPlum.transfection control The "Damaged 2 cocktail" contained irradiated plasmid (BFP), GFP transfection control, T:G mismatch (mOrange), and dA opposite 8oxodG (mPlum). Removal of the lesion turns off fluorescence intensity for uracil, hypoxanthine, and both 8oxodG plasmids. Repair of T opposite G turns on fluorescence. Daily performance tests on the flow cytometer using the manufacturer's performance tracking beads.