Screening for inhibitors of RAD52-ssDNA exerting synthetic lethality in BRCA-deficient cells.

<p>(<b>A</b>) The indicated compounds, F79 aptamer (F79), or DMSO (-), were added to the mixture of GST-RAD52 and IRDye800-ssDNA (ssDNA = IRDye800-ssDNA only). RAD52-ssDNA complex, free ssDNA, and RAD52 protein were detected by fluorescence and Western analysis, respectively. Compound 46611 was used as negative control. Results represent at least 3 experiments. (<b>B</b>) Growth of BRCA1-null HCC1937 cells (grey bars) and their BRCA1-reconstitued counterparts (black bars) in the presence of diluent, 5 μM F79 and indicated compounds (10 μM of 63878 and 20 μM of the others). Number of cells ± SD normalized to untreated controls from 3 experiments in triplicates, *p<0.01. (<b>C</b>) Growth of BRCA1-null HCC1937 cells (grey dots) and their BRCA1-reconstitued counterparts (black dots) in the presence of indicated concentrations of AICAR. Number of cells ± SD normalized to untreated controls from 2 experiments in triplicates. (<b>D</b>) Growth of BRCA2 mutated human pancreatic adenocarcinoma Capan-1 cells (grey bar) and their BRCA2 wild-type reconstituted counterparts (black bar) in the presence of 20 μM AICAR. Number of cells ± SD normalized to untreated controls from 2 experiments in triplicates, *p<0.03.