Fig. 1

QA impairs mitolysosome formation, which is restored by mitophagy inducer UA in microglia. (A) Schematic working model of Cox8-EGFP-mCherry mitophagy reporter construct. Created with BioRender.com (B) Representative confocal images of BV2 cells transiently transfected with Cox8-EGFP-mCherry plasmid treated with or without 50 µM QA for 72 h. Scale bar, 5 μm. (D) Representative confocal images of HMC3 cells transiently transfected with COX8-EGFP-mCherry plasmid treated with or without 50 µM QA for 24 h in the absence or presence of 10 µM UA. Scale bar, 10 μm. (F) Representative confocal images of 50 µM QA-treated HMC3 cells transiently transfected with Cox8-EGFP-mCherry plasmid in the absence or presence of 10 µM UA and/or 20 µM CQ for 24 h. Scale bar, 10 μm. (C, E, and G) Graphs represent the number of mitolysosomes (red dots) per cell in indicated conditions. Data are means ± SEM. (C) n = 12–15 cells, (E) n = 10 cells, and (G) n = 8 cells; *** p < 0.001 versus no treatment (control) group (C, E), *** p < 0.001 versus QA alone treatment group (G), ^###p < 0.001 versus QA alone treatment group (E), ^###p < 0.001 versus QA and UA treatment group (G); Two-tailed unpaired t test (C) or one-way ANOVA followed by Tukey’s multiple comparisons test (E and G)