Fig. 5

Oxidative stress impairs p62 degradation in microglia. (A) Representative western blot images of LC3B-I/II and p62 in BV2 microglia treated with or without 150 µM H₂O₂ for 72 h. β-actin was used for loading control. (B) Quantification of relative protein level of LC3B-II in A (n = 3). (C) Quantification of relative protein level of p62 in A (n = 5). (D) Representative confocal images of BV2 cells transiently transfected with Cox8-EGFP-mCherry plasmid with or without 150 µM H₂O₂ treatment for 72 h. Scale bar, 5 μm. (E) Graph represents the number of mitolysosomes (red dots) per cell in D (n = 18 cells). (F) Representative western blot image and quantification of relative protein level of p62 in BV2 microglia treated with or without 150 µM H₂O₂ in the absence or presence of UA as indicated for 72 h. β-actin was used for loading control (n = 3–4). (G) Representative western blot image and quantification of relative protein level of p62 in BV2 microglia treated with or without 10 µM UA for 72 h. β-actin was used for loading control (n = 6). Data are means ± SEM. *p < 0.05, **P < 0.01 and ***P < 0.001 versus no treatment (control) group, ^#p < 0.05 and ^##p < 0.01 versus H₂O₂ alone treatment group; Two-tailed unpaired t test (B, C, E and G) or one-way ANOVA followed by Tukey’s multiple comparisons test (F)