Effects of RU.521 on eNOS, cell senescence, inflammatory cytokines and NO production in D-GAL-treated HAECs. 3μM RU.521 was used to treat the HAECs. eNOS, p53, p21, p16, cGAS, p-STING, STING, IRF3, p-IRF3 and β-actin expression levels in HAECs from the Control, D-GAL and D-GAL+RU.521 groups were examined by western blot analysis (A). The β-actin was used as the housekeeper protein for normalization. Quantification of the protein levels is shown in (B). mRNA expression of IFNβ (C), Ifit1 (D), Ifit2 (E), Ifit3 (F), MCP-1 (G), IL-1β (H), IL-6 (I) and TNF-α (J) in HAECs from the Control, D-GAL and D-GAL+RU.521 groups was determined by PCR. NO release into the culture medium of HAECs from the Control, D-GAL and D-GAL+RU.521 groups was measured (K). Representative photomicrographs and quantitative analysis of SA-β-gal-positive staining in the Control, D-GAL and D-GAL+RU.521 groups (L and M). Data were analyzed by one way ANOVA plus Bonferroni post hoc test. All data shown are mean±SD. AU indicates arbitrary units. Relative expression is the fold changes relative to the Control group. n=6, *P<0.05 compared with the Control group, #P<0.05 compared with the D-GAL group.