Fig. 9.

The mutation of residue Arg483 in KEAP1 alleviates the anti-oxidative stress and anti-pyroptosis effect of quercetin pretreatment.

The plasmid with KEAP1-WT, single mutation at R415S or R483S are transfected into macrophages, quercetin (50 μM) is used to pre-treat macrophages, followed by ox-LDL (100 μg/ml) combined in different groups. (A). Western blot is used to evaluate the KEAP1 level in different groups (n = 3, *p < 0.05 vs NC group), Analyzed by Student's t-test. (B). DCFH-DA staining is used to display the ROS level in macrophages (n = 3), Analyzed by Student's t-test. (C–F). Hoechst/PI staining (n = 5), CCK-8 (n = 7) and LDH assay (n = 4) are used to measure the cell viability of macrophages (n = 5). Analyzed by Mann-Whitney U test. (G) The MDA level in macrophages (n = 8). Analyzed by Mann-Whitney U test. (H) The SOD activity in macrophages (n = 5). *p < 0.05, **p < 0.01, ***p < 0.001, vs ox-LDL group.