Fig. 5

GEVs regulated proinflammatory cytokine transcription and secretion through NF-κB signaling pathways in a positive feedback manner. Mouse macrophages were pretreated with the IκBα phosphorylation inhibitor BAY 11-7082 (5 μM) for 2 h. Groups not pretreated were used as controls. The cells were inoculated with GEVs for 0.5 h. a The protein expression levels of phosphorylated phospho-IκBα were measured by western blotting, and gray values of phosphorylated protein/β-actin were calculated using ImageJ software. b–d Transcription levels of proinflammatory cytokines in cells collected 6 h after inoculation were determined using qPCR assays. e–g Transcription levels of proinflammatory cytokines in cells collected 12 h after inoculation were determined using qPCR assays. h–j The proinflammatory cytokine protein secretion levels in the supernatants collected 18 h after inoculation were detected using ELISAs. The results are shown as the mean ± SEM of triplicate experiments. Significant differences vs noninhibitor treatment control: **p < 0.01 , ***p < 0.001. ND Not determined