Inhibition of uracil removal by ATA in MCF7 cells

Testing the inhibition of uracil removal from DNA by the inhibitor ATA. Conclusion: ATA inhibits the removal of uracil from a plasmid based assay in MCF7 cells Notes: MCF7 cells were seeded into plates and treated with DMSO or with a putative uracil dna glycosylase inhibitor (ATA, sometimes referred to as ATA mix). Following treatment, cells were transfected with a plasmid-based reporter that measures uracil removal. The samples labelled "Undamaged" were transfected with plasmids encoding BFP and GFP. The samples labeled "Damaged" were transfected with a plasmid encoding GFP and a uracil-containing plasmid encoding BFP. If uracil is present in the plasmid, wild type BFP transcripts are produced. If uracil is removed. non-fluorescent transcripts are produced. Hence, increased BFP fluorescence in the samples labeled "Damaged" means decreased removal of uracil from DNA. Daily performance tests on the flow cytometer using the manufacturer's performance tracking beads.