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. 2011 Jun 3;12(7):713–719. doi: 10.1038/embor.2011.96

Figure 5.

Figure 5

Human polynucleotide kinase/phosphatase is recruited to DNA sites damaged within seconds after damage induction. (A) Rapid recruitment of dsRED–PNKP to DNA sites damaged by a focused laser microbeam. (B) Human PNKP recruited to the damaged site and co-localized with 53BP1, a central factor in the early phase of the DNA-damage response. Scale bars, 10 μM. (C) Recruitment kinetics of dsRED-tagged human PNKP in wild type (WT) and phospho-mutants during the first 3 h after DNA damage induction. The cell nuclei were exposed to a single burst of laser irradiation, images were captured at the indicated time the points and the amounts of dsRED–PNKP were quantified (Yano et al, 2008; Moyal et al, 2011). To compensate for nonspecific photobleaching, the background fluorescence was subtracted from the accumulation spot. Each data point is the average of 10 independent measurements. Error bars represent s.d. PNKP, polynucleotide kinase/phosphatase.