Fig. 3

Analysis of the effects of DMF on p38, JNK, Akt and p42/44 phosphorylation in TNF-α-treated HUVECs and the influence on MCP-1 expression. a p38, JNK, Akt and p42/44 expression and phosphorylation: Western blot analysis of HUVECs treated with vehicle (solvent only), TNF-α (20 ng/ml) or DMF (80 μM, 3 h pre-treatment) + TNFα for 60 min. b Densitometry analysis: The results were normalized to the expression of the nonposphorylated controls (p38, JNK, AKT, p42/44). The relative expression of the posphorylated protein is presented in arbitrary units (arb. units). The mean values from at least three independent experiments are presented as the mean ± standard deviation. The TNF-α only treated bands served as control to the combinational treatment (DMF+ TNF-α). We analyzed theata using the Student’s t test. *p <0.05; n.s. not significant. c Signaling pathway blockade: MCP-1 ELISA 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. We performed the blockade of the signaling pathways via treatment with SB203580 (1 μM), Wortmannin (5 μM) and PD 98059 (30 μM) 30 min before the main treatment. We present the mean values from three triplicate experiments as the mean ± SEM. We analyzed the data using the Student’s t-test. *p < 0.05 versus TNF-α; **p < 0.05 versus control