Fig. 5

The regulatory effect of GDNPs on the Keap1–Nrf2–p62 signaling pathway in an in vitro model. Protein expression of GCLC, GCLM, HO-1, Nrf2, Keap1, p62, Phospho-sequestosome 1 (p-p62) and β-actin measured by western blotting in RAW264.7 cells treated with and without different concentrations of GDNPs (A). Western blot bands demonstrating Nrf2 translocation from cytoplasm to nucleus. B Quantitative plot of Nrf2 fluorescence expression (C). Nrf2 fluorescence expression in RAW264.7 cells. Scale bar: 400 μm (D). Protein expression of HO-1, Nrf2, NQO1 and β-actin measured by western blotting under the action of the inhibitor ML385 in in RAW264.7 cells. E Levels of IL-6 in cell supernatants in the presence or absence of the ML385 inhibitor (F). Image processing and densitometric analysis were performed using ImageJ analysis software. Data are expressed as mean ± SD. n = 3. ^#p < 0.05 and ^##p < 0.01 vs. Control; *p < 0.05 and **p < 0.01 vs. LPS-stimulated cells (One-way ANOVA and Dunnett’s post-hoc test)