Figure 5
3-MA inhibits cytokines-induced NF-κB p65 nuclear translocation. (A) HUVECs were pretreated with or without 10 μmol/L Bay11 for 30 min followed by 10 ng/mL IL-1β or 10 ng/mL TNFα for 24 h. VCAM-1 and ICAM-1 were analyzed by western blotting. Upper panel shows a representative blot (n = 3) and the lower panel the quantitative analysis. *Indicates P < 0.05. (B) HUVECs were pretreated with or without 1 μmol/L BI605906 or 100 nmol/L NG25 for 30 min followed by 10 ng/mL IL-1β or 10 ng/mL TNFα for 24 h. VCAM-1 and ICAM-1 were analyzed by western blotting. (C,D) HUVECs were treated with 10 ng/mL TNFα (C) or 10 ng/mL IL-1β (D) for indicated time. The nuclear fraction (Nuc) and cytoplasmic fraction (Cyto) were separated. p65 and p50 in each fraction were analyzed by western blotting. (E,F) HUVECs were pretreated with 5 mmol/L 3-MA, 10 μmol/L MG-132, 10 μmol/L Bay11, or 1 μmol/L BI605906 for 30 min followed by 10 ng/mL TNFα (E) or 10 ng/mL IL-1β (F) for 6 h. p65 in nuclear fraction was analyzed by western blotting. Upper panel shows a representative blot and the lower panel the quantitative analysis. Each bar denotes mean ± SD of three independent experiments. *Indicates P < 0.05.
