Fig. 3.

ZNF281 recruitment to DNA lesions depends on PARP. a, b U2OS cells stably expressing EGFP–ZNF281 were treated with siRNAs targeting XRCC4 (a), Ku86 (b) or DNA-PK-cs (Supplementary Fig. S3d). ZNF281 relocalisation to the UV-induced DNA lesions in silenced cells was measured and compared to siCtrl-treated cells. Fifty to 75 cells were analysed at each time point in three independent experiments. Graphs present means ± SEM. siCtrl-treated cells were plotted separately against individual siRNA-treated cells to clearly present the data. c U2OS–EGFP–ZNF281 cells were pre-treated with 10 μM PARPi (Olaparib) for 1 h before laser microirradiation. Data were obtained from 60 to 75 cells in three independent experiments. Graphs present means ± SEM. d Recruitment of ZNF281 wild-type (Wt) and ZNF281 4X Zinc mutant (4X C>A) to UV laser-induced DNA damage sites in U2OS cells. Eighty cells expressing the ZNF281 4X C>A mutant were analysed from three independent experiments. Mean values ± SEM are plotted against the ZNF281 Wt curve (from Fig. 1a) for comparison. Representative images acquired before and after damage are shown below the graphs of the quantitative analysis in panels c and d. White circles indicate the irradiated area. Scale bars, 10 μm