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. 2022 Apr 15;12:6335. doi: 10.1038/s41598-022-10018-0

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© The Author(s) 2022

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Detection of DNA damage in CD4 + T, CD8 + T, B cells and monocytes using HTI. (a) CD4 + T cells were either left untreated or treated with 30 µM ETP for 1.5 h before being subjected to HTI. DNA damage sites are indicated by formation of 53BP1 and γ-H2AX foci detected by specific antibodies. Scale bar: 10 µm. (b) Distribution of 53BP1 sum spot intensity in DMSO- or ETP-treated cells. ****: p < 0.0001. (c) Distribution of γ-H2AX mean nuclear intensity in DMSO- or ETP-treated immune cells. ****: p < 0.0001 using Mann–Whitney-Wilcoxon test. Number of analyzed cells (n) indicated for each sample. The values are representative of at least 3 experiments. Boxes represent the first and third quartiles of the distribution. The line represents the median. The whiskers represent the highest and lowest data points within 1.5 times the interquartile range above the upper quartile and below the lower quartile. (d) Two-dimensional kernel density contour plots of the correlation between 53BP1 sum spot intensity and γ-H2AX mean nuclear intensity for both ETP-treated (blue) and untreated (red) immune cell lines for the dataset shown in panels B and C. N = 3 wells per sample with at least 185 cells per well.