Fig. 4

Analysis of TNF-α-induced p65 nuclear entry, phosphorylation (Ser 536), promoter activity and IκBα degradation during DMF treatment. a Nuclear p65 translocation and phosphorylation: Western blot analysis of nuclear proteins of HUVECs treated with vehicle (solvent only), TNF-α (20 ng/ml) or DMF (80 μM, 3-h pre-treatment) + TNF-α for 60 min. The phosphorylated p65 (Serine 536) band is marked by an arrow head. b Representative immunofluorescent analysis of p65 in HUVECs that were treated with vehicle, TNF-α (20 ng/ml) or DMF (80 μM, 3 h pre-treatment) + TNF-α for 1 h. c IkB degradation: Western blot analysis cytosolic proteins of HUVECs treated with vehicle (solvent only), TNF-α (20 ng/ml) or DMF (80 μM, 3 h pre-treatment) + TNF-α for 60 min. d Analyses of the NfκB luciferase (Luc) reporter constructs in HUVECs treated with vehicle (solvent only), DMF (80 μM), TNF-α (20 ng/ml) or DMF (80 μM) + TNF-α for 24 h, respectively. The Luc activities are expressed as relative luciferase activity as a percent (mean ± SEM of at least five independent triplicate assays). *p < 0.05 versus TNF-α; **p < 0.05 versus control