Journal: Cancers

Article Title: Characterisation of Ovarian Cancer Cell Line NIH-OVCAR3 and Implications of Genomic, Transcriptomic, Proteomic and Functional DNA Damage Response Biomarkers for Therapeutic Targeting

doi: 10.3390/cancers12071939

Figure Lengend Snippet: Assessment of HRR and NHEJ in the NIH-OVCAR3 cells. ( A ) Cells were exposed to 0.5% DMSO (control) or 10 µM rucaparib for 48 h before being fixed and stained as per the protocol outlined in the methods. After imaging, the number of γH2AX and RAD51 foci per nucleus was calculated using ImageJ software, data points represent number of foci in an individual nucleus. The fold change in number of foci per nuclei between control and 10 µM rucaparib was then calculated. ( B ) Cells were co-transfected with pDRGFP and pCBASceI plasmids and analysed for the presence of GFP-positive cells 48 h post-transfection. % HRR activity was calculated by normalizing to the untransfected control. Graph shows mean of 2 individual experiments. ( C ) Cells were co-transfected with pimEJ5GFP and pCBASceI plasmids and analysed for the presence of GFP-positive cells 48 h post-transfection. % NHEJ activity was calculated by normalizing to the untransfected control. Graph shows mean of two individual experiments. [* p < 0.05; ** p < 0.005; **** p < 0.0001 (unpaired t -test)].

Article Snippet: pDRGFP (plasmid # 26475, Addgene, Watertown, MA, USA) and pCBASceI (plasmid # 26477, Addgene) plasmids used for the HRR assay were a kind gift from Maria Jasin [ , ].

Techniques: Control, Staining, Imaging, Software, Transfection, Activity Assay