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. 2022 Aug 1;14(5):1077–1101. doi: 10.1016/j.jcmgh.2022.07.014

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© 2022 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

FGL2 promoted NETs formation by induction of autophagy after MHV-3 challenge. LC3, P62, Beclin 1 (left panel), PAD4, and Cit-H3 (right panel) expression levels in (A) hepatic tissues and (B) MHV-3–stimulated BM neutrophils were detected by Western blot and quantified by Image Lab (n = 5 in each group). (C) Autophagosome formation was detected by fluorescent staining of LC3 in the presence of autophagy inhibitor (3-MA) or not. Scale bars: 10 μm. The numbers of LC3 particles were counted in 3–5 microscopic fields per slide from 3 slides in each group. (D) Transmission electron microscopy images showed the presence of double-membrane autophagosomes (arrowheads) in isolated neutrophils. Scale bars: 500 nm. (E) BM neutrophils were treated with MHV-3 in the presence of 3-MA or not. The induction of NETs was measured by fluorescent microscopy using Sytox Green. (F) MPO-DNA levels in the supernatant were detected. n = 3 in each group. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001, determined by 1-way analysis of variance. DAPI, 4′,6-diamidino-2-phenylindole; OD, optical density.