Fig. 4

Efficient processing of DNA lesions is necessary to prevent cell cycle exit. a Schematic representation of the experimental set-up. b Stills represent RPE CCNB1^YFP cells with 53BP1^mCherry tracked by live-cell imaging at indicated timepoints after IR (2 Gy) followed by an image of the identical cell that was fixed 4 h after IR and stained for RPA and Rad51. Scale bar 10 μm. c Number of indicated foci in cells that have cytoplasmic Cyclin B1 localization at 4 h after IR (2 Gy) or cells that translocated Cyclin B1 and therefore have nuclear localization at 4 h after IR. 53BP1 foci were determined at indicated timepoints by live-cell imaging of RPE CCNB1^YFP-53BP1^mCherry cells. RPA and Rad51 foci were determined in the tracked cell by imaging of the same position after fixation, pre-extraction and antibody staining for RPA and Rad51. Pooled cells from three independent experiments. n > 65 + sd