Journal: Nature communications

Article Title: Genome-wide Transcriptome Profiling of Homologous Recombination DNA Repair

doi: 10.1038/ncomms4361

Figure Lengend Snippet: ( a ) Modified HR repair assay was performed by transfecting cells with DRGFP DSB substrate plasmid and I-SceI plasmid through electroporation at 270V, 975uF using a BioRad genepulsar II. Flow cytometry analysis was performed 48 to 72 hours later to detect GFP-positive cells. Each value is shown as mean + SD for three independent experiments. Student’s t -test showed increased HR repair efficiency in HR-intact cell lines compared to the HR-deficient cell lines in each cancer type, respectively. ( b , c ) Colony formation assay was performed with indicated concentrations of olaparib ( b ) or rucaparib ( c ). Each value is relative to the value in the cells treated with vehicle control. Results are shown as mean + SD from three independent experiments. Student’s t -test showed that the drug response to PARP inhibitors differed between cancer cell lines with and without the HRD gene signature (P<0.05 through panel b to c).

Article Snippet: For cell lines that do not stably contain the DR-GFP plasmid, 1×10 6 cells were electroporated with 12 μg of DR-GFP and 12 μg of pCBASce plasmids at 270V, 975uF using a BioRad genepulsar II .

Techniques: Modification, Plasmid Preparation, Electroporation, Flow Cytometry, Colony Assay