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. 2022 Jan 24;13:816037. doi: 10.3389/fimmu.2022.816037

Figure 3.

Figure 3

The evaluation of function of Neu during 6 days in 2D and 3D culture system. (A) Flow cytometry analysis of ROS produced by mouse peripheral blood Neu cultured in 2D and 3D systems and corresponding statistical results (B). Chemotactic ability of mouse peripheral blood Neu shown as the number of cells per high power field on the Petri dish bottom in 2D culture and 3D culture systems for control (randomly migrating) and WKYMVm (100 nM) stimulated mouse peripheral blood Neu (C) or control and fMLF (1 μM) stimulated human peripheral blood Neu (D). (E) Human Neu released NETs marked by NE (Alexa Fluor 594 dye, red) and nuclear (SYTO 13, green) when stimulated by 20 μM PMA for 4 h in 2D and 3D culture systems under a confocal microscope in vitro. (F) A significantly different dsDNA release was detected using fluorescent-based Picogreen assay. (G) Meanwhile, a significantly different release of neutrophil elastase coupled dsDNA was measured using fluorescent-based Picogreen assay coated with NE antibodies. Error bars represent mean ± S.D. n = 3, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns, not statistically significantly different by Two way ANOVA.