Fig. 4From: Extracellular vesicles produced by avian pathogenic Escherichia coli (APEC) activate macrophage proinflammatory response and neutrophil extracellular trap (NET) formation through TLR4 signalingLoss of LPS significantly weakened the inflammatory response in THP-1 macrophages. (A + B) Escherichia coli CT265, mutant strain CT265ΔwecA, and EVs from both bacteria were analyzed with the PyroGene™ Recombinant Factor C Endotoxin Detection Assay. Statistical test: one-way ANOVA, *P < 0.05, **P < 0.01. (C) TLR4 mRNA levels in PMA-treated THP-1 macrophages, pretreated or not pretreated with TAK242 and incubated for 12 h with EVs lacking LPS (30 µg/mL), were measured with RT–qPCR. Statistical test: one-way ANOVA, *P < 0.05, **P < 0.01. (D) Expression levels of inflammasome-related proteins in PMA-treated THP-1 macrophages incubated for 12 h with CT265ΔwecA EVs (30, 40, or 50 µg/mL) were measured with western blotting assays. A representative western blot is shown. (E) Densitometric analysis was used to quantify the levels of inflammasome-related proteins. Statistical test: two-way ANOVA, *P < 0.05, **P < 0.01Back to article page