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. 2020 May 7;10:7734. doi: 10.1038/s41598-020-64672-3

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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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Fractionated, compared to single exposure, radiation profoundly suppresses levels of KLF2 and KLF4. Representative (3–5 independent experiments) Western blot analysis and quantification of KLF2 (n = 5) and KLF4 (n = 3) levels in whole-cell lysates from nonirradiated (sham) and irradiated HUVECs 4 h and 24 h after exposure to (a) either five fractions of 2 Gy (5 × 2 Gy) or single exposure to 10 Gy and (b) either five fractions of 2.5 Gy (5 × 2.5 Gy) or single exposure to 12.5 Gy. Fractions delivered at 24-h intervals. β-actin served as a loading control. c Representative photomicrograph (20× magnification) showing immunofluorescence of KLF2 (red) in nonirradiated (sham) and irradiated HUVECs 4 h after exposure to either five fractions of 2 Gy or single exposure to 10 Gy (n = 3). Nuclei were stained with DAPI (blue). d Representative Western blot analysis and quantification of KLF2 (n = 3) and γ-H2AX (n = 3) phosphorylation levels in nonirradiated (0 h) and irradiated HUVECs at indicated time intervals after single exposure to 2 Gy and 5 Gy. β-actin served as a loading control. (n, number of independent experiments performed; a, significant statistical difference between nonirradiated and irradiated groups; b, significant statistical difference between fractionated irradiation and single exposure; *, p < 0.05; **, p < 0.01; ***, p < 0.001).