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. 2021 Nov 2;11:21439. doi: 10.1038/s41598-021-00863-w

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© The Author(s) 2021

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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Damage to endothelial cells by NETs. To assess HUVEC damage, control (0 nM PMA), 400 nM PMA, HV serum, and HD serum were applied to dHL-60 cells for 17 h. Supernatants (10%) were added to HUVECs and incubated for 20 h. Serum-free medium (SFM) and 10% FBS were used as controls. (A) Morphology of HUVECs. In HV and HD NETs, cells became elongated compared to the control, and many dead floating cells were observed (magnification, upper panels, ×100; lower panels, ×400). (B) Effect of NETs from dHL-60 cells on HUVEC viability was tested by WST-8 assay. HUVEC viability was higher in HV serum-induced NETs than in uremic serum-induced NETs and SFM, and lower than those in 10% FBS. (C) Repeat experiments with isolated neutrophils from blood donor. Results are means ± SEM, Student’s t-test, ***p < 0.001.