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. 2021 May 26;12:629167. doi: 10.3389/fimmu.2021.629167

Figure 1.

Figure 1

Circulating NETs in dengue patients are mainly NOX-independently generated. (A) Time course of total NETs (μg/mL) in plasma of dengue patients according to day since fever onset and in a group of healthy controls (HC). Depicted are individual data together with a box plot showing median with interquartile range. Differences in the dengue patients over time were analyzed using the Wilcoxon signed rank test; differences with the HC group were analyzed using Mann-Whitney U test; *p < 0.05, **p < 0.01, ***p < 0.001. (B) NET formation (ng/ml) after incubation of neutrophils with 10% plasma of dengue patients according to day since fever onset and of a group of HC. Depicted are individual data of five plasmas per group tested on 3 neutrophil donors, together with a box plot showing median with interquartile range. Differences with the HC group were analyzed using Mann-Whitney U test;, ***p < 0.001. (C) Scatterplot quadrant analysis to differentiate NOX-dependent and NOX-independent NET formation on the basis of results of the total NETs (X-axis) and MPO-H4K8Ac, K12Ac, K16Ac assay (Y-axis). Cut-off points were determined from mean values and standard deviations from HC samples (orange and blue-dotted lines). Samples positive to both assays were classified as NETs derived NOX-independently, while samples only positive for MPO-DNA were classified as NETs derived from NOX-dependently. HC, healthy controls; NOX, NADPH-oxidase.