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. 2020 Jan 29;10:1443. doi: 10.1038/s41598-020-58084-6

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Live cell imaging of real time NBS1 focus formation using X-rays. (A) Selected images of a movie showing NBS1-GFP foci formation during radiation with sparsely ionising radiation. (B) Nuclear wide analysis of recruitment kinetics showing nearly identical NBS1 recruitment kinetics for X-ray induced (n = 25 up to 10 min and 4–14 for the longer time points) and delta-ray induced DSBs (n = 73; Error bars 95% CI). Data were normalised to their maximum at around 20 min. (C) Analysis of NBS1 recruitment kinetics to individual X-ray induced DSBs showing a fraction of clearly delayed recruiting foci. (D) Distribution of lag-phases showing similarity after X-rays (n = 333) to the lag phase distribution of off-track- foci after HZE particle irradiation (from Fig. 2C; n = 189). Image montage of (A) was done using ImageJ 1.48 v (https://imagej.nih.gov). Graphs were generated using Origin Pro V.2019 (Originlab Corp., Northampton, MA, USA).