Fig. 5: IL-1β mediates the macrophages inducing NETosis under poly(I:C) stimulation.
From: Hippo pathway and NLRP3-driven NETosis in macrophages: Mechanisms of viral pneumoniaaggravation
A Relative expression of IL-1β protein in RAW246.7 cells treated with poly(I:C) and Hippo pathway inhibitors was assessed. B Quantitative analysis of the protein IL-1β relative to Tubulin under different conditions. C Measurement of IL-1β level in culture supernatants from RAW246.7 cells treated with poly(I:C) and Hippo pathway inhibitors Lats-IN-1 by ELISA. D NETosis biomarker levels in the cocultures of neutrophils and conditioned medium for RAW246.7 cells under poly(I:C) stimulation and neutralized IL-1β antibody were assessed by immunofluorescence. E Graphic presentations of fluorescence mean densities of CitH3 and MPO under different conditions. F Evaluation of CitH3 protein expression in the co-cultures of neutrophils and conditioned medium for RAW246.7 cells under poly(I:C) stimulation and neutralized IL-1β antibody by western blotting. G Quantitative analysis of the protein CitH3 relative to Tubulin. H dsDNA and LL-37 levels in the cocultures of neutrophils and conditioned medium for RAW246.7 cells under poly(I:C) stimulation and neutralized IL-1β antibody were evaluated by ELISA. Note: IN, Lats-IN-1 is a potent and ATP-competitive inhibitor of LATS1 and LATS2 kinases. All data are representative as means ± s.e.m of three independent experiments. Student’s t test for (A–H); *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. Scale bar = 20 μm.
