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. 2014 Mar 3;42(8):4996–5006. doi: 10.1093/nar/gku160

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© The Author(s) 2014. Published by Oxford University Press.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 5. — Artemis is required for the DNA-repair capability in K-H deficient cells. (A) Basal levels of the DNA-damage repair biomarker 53BP1 were measured in shScr, shk-h and shk-h cells stably over-expressing Artemis (Art), (shk-h + Art), by immunofluorescence (IF). (B) Sensitivity to IR in shScr, shk-h and shk-h + Art, mock untreated or IR exposed cells was monitored by colony forming assay. (C) Disappearance of 53BP1 foci was measured in shScr, shk-h and shk-h + Art at indicated times after IR, first time point measured is 0.5 h after IR exposure. (D) The ability to perform NHEJ in shScr, shk-h and shk-h + Art was monitored by a plasmid-based NHEJ assay with plasmids digested with either HindIII to study compatible end re-ligation, or I-SceI to study incompatible end re-ligation. (E) Basal levels of 53BP1 foci were measured in wild type (mk-h^+/+), K-H heterozygote (mk-h^+/–) and Artemis deficient (mart^–/–) MEFs by IF. (F) Rate of 53BP1 foci disappearance after IR at times indicated was measured in mk-h^+/+, mk-h^+/– and mart^–/– MEFs by IF, first time point is 0.5 h after IR. Cells (300) were visualized for 53BP1 foci. Colonies were determined as ≥50 normal-appearing cells in a 7-day period. Events (10 000) were counted by flow cytometry. (**P < 0.01, *P < 0.05).