IL20RB Antibody - #DF3603

Figure 7. Interleukin-19 (IL-19) induces neutrophil extracellular trap (NET) formation in vitro through IL-20Rβ receptor signaling. (A) Schematic of Transwell migration assay for neutrophils. (B) Neutrophils were isolated from mouse bone marrow and loaded into the upper Transwell chamber. Serum-free RPMI-1640 medium (500 μL) containing 1% penicillin/streptomycin (P/S), with or without different concentrations of IL-19, was added to the lower chamber. Neutrophil migration was detected by crystal violet staining. Neutrophils are marked by black arrows (n = 3). Scale bar, 100 μm. (C) Mouse bone marrow-derived neutrophils were treated with different concentrations of IL-19 (5 ng/mL, 20 ng/mL, 80 ng/mL) for 4 h and then collected. (D and E) Level of myeloperoxidase (MPO) and dsDNA in the cell culture supernatant assessed after treatment with different concentrations of IL-19 (n = 3). (F) Level of nucleosomes in the cell culture supernatant assessed after treatment with different concentrations of IL-19 (n = 3). (G) Protein levels of IL-20Rβ and citrullinated histone 3 (Cit-H3) assessed by western blotting after treatment with different concentrations of IL-19 (n = 5). (H and I) Immunofluorescence staining of IL-20Rβ and Cit-H3 of neutrophils after treatment with different concentrations of IL-19 (n = 3). Scale bar, 100 μm. (J) Representative scanning electron microscopy images of vehicle- and IL-19-stimulated neutrophils (NETs, n = 3). Scale bar, 5 μm. Data are presented as the mean ± SEM of three biological replicates per condition. Each dot represents a sample. Statistically significant differences were determined by an independent sample t-test and one-way ANOVA followed by Tukey’s post hoc test (D–G). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns: non-significant. Figure 7—source data 1 Figure 7—source data 2 Interleukin-19 (IL-19) induces neutrophil extracellular trap (NET) formation in vitro through IL-20Rβ receptor signaling.