Fig. 3

CoQ10 inhibits the ROS generation and activation of NLRP3/IL1β signaling in macrophages. (A) Gene expression analysis in peritoneal macrophages with or without 6-hour stimulation of LPS/INFγ, in the presence or absence of CoQ10 (Statistical analysis was performed using 2-way ANOVA with Sidak’s multiple comparison tests). (B) Heatmap of RNA script reads of specific genes related to inflammatory cytokines, inflammatory chemokines, macrophage activation, NOD-like receptor signaling, coenzyme metabolic process, fatty acid beta oxidation, ROS biosynthetic process between LPS/IFNγ-primed macrophages treated with or without CoQ10. (C) KEGG enrichment analysis of top-ranked pathways of differentially expressed genes between LPS/IFNγ-primed macrophages treated with or without CoQ10. (D) Western blot analysis of intracellular NLRP3 in peritoneal macrophages after 6 h of stimulation with LPS/INFγ in the presence of vehicle or CoQ10 (Statistical analysis was performed using a two-tailed unpaired Student’s t-test). Data were normalized to β-actin. (E) Western blot analysis of intracellular pro-IL1β in the macrophages after 6 h of stimulation with LPS/INFγ in the presence of vehicle or CoQ10 (Statistical analysis was performed using a two-tailed unpaired Student’s t-test). Data were normalized to β-actin. (F) Western blot analysis of intracellular cleaved-IL-1β in the macrophages after 6 h of stimulation with LPS/INFγ combined with a final incubation with 5mM ATP in the presence of vehicle or CoQ10 (Statistical analysis was performed using a two-tailed unpaired Student’s t-test). Data were normalized to β-actin. (G) ELISA analysis of secreted IL-1β levels in the supernatant of cultured macrophages after 6 hours’ LPS/INFγ stimulation combined with a final 30 min incubation with 5mM ATP in the presence of vehicle or CoQ10 (Statistical analysis was performed using a two-tailed unpaired Student’s t-test). LPS: lipopolysaccharide; IFNγ: interferon γ; ATP: adenosine triphosphate; CoQ10: Coenzyme Q10; ROS: reactive oxygen species. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001