Fig. 6.

Inhibiting autophagy reverses the protective effect of Qu on LPS + ATP-induced mitochondrial injury and inflammation in BV2 cells.

BV2 cells were treated with 3-MA (5

mM, 1 h

) before Qu (30 μM, 1 h) treatment followed by LPS (100 ng/ml, 24 h) and ATP (5 mM, 30 min) stimulation.

(a–b) Mitochondrial ROS levels were assessed by staining with MitoSOX and analyzed using confocal microscopy (a). DAPI was used to stain nuclei (blue). Scale bars, 40 μm. Quantification of MitoSOX fluorescence intensity using ImageJ software (b).

(c–d) Mitochondrial morphology in BV2 cells were monitored and pictured by immunostaining for Tomm20. Scale bars, 20 μm.

(e) ELISA of IL-1β in supernatants from differently treated BV2 cells.

(f–g) Expression of pro-IL-1β in the cell lysate (Ly) and cleaved IL-1β in the supernatant (SN) were detected by an immunoblot assay (f) and quantified as normalized to the control group (g). Data are shown as the mean ± SEM from three to five independent experiments. *P < 0.05, **P < 0.01 and ***P < 0.001 by one-way ANOVA followed by Tukey's post hoc test. Ctl, untreated control; Qu, quercetin; LPS, lipopolysaccharide; 3-MA, 3-methyladenine. . (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)