Coenzyme Q₀ Inhibits NLRP3 Inflammasome Activation through Mitophagy Induction in LPS/ATP-Stimulated Macrophages

Figure 3

CoQ₀ induces autophagy in LPS/ATP-stimulated RAW264.7 macrophages. Cells were first treated with 2.5-10 μM of CoQ₀ and/or 2.5 mM 3-MA (autophagy inhibitor) for 1 h and then followed by stimulation of LPS (1 μg/mL) for 5 h and ATP (5 mM) for 1 h. (a) The expression of LC3-I/LC3-II and p62/SQSTM1 protein was estimated by Western blot. (b) The modifications in LC3B expression were observed by immunofluorescence staining. Cells were incubated with anti-LC3B antibody followed by secondary antibody labelled with FITC. 630x magnification of a confocal microscope was used to visualize the subcellular localization of LC3B. (c) Fold changes in LC3B were determined. (d) CoQ₀ affects Beclin-1 and Bcl-2 expression in a dose-dependent manner shown by Western blot analysis. (e) Beclin-1/Bcl-2 ratio relative changes were measured by commercial software representing control as 1-fold. The results were calculated as the of three independent experiments. , compared with untreated control cells, and ^#; ^##; ^### compared with 3-MA- or LPS/ATP-stimulated cells assigned statistically significant.

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