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. 2013 Sep 2;8(9):e73122. doi: 10.1371/journal.pone.0073122

Figure 1. Influence of NOD on the expression of adhesion molecules and HO-1.

Figure 1

(A) HUVECs were stimulated for 24 hrs with 10 ng/ml of TNF-α in the presence of different concentrations of NOD. The expression of VCAM-1 and HO-1 was assessed by western blotting. GAPDH was used as loading control. (B) HUVECs were stimulated as described in A. The expression of ICAM-1 and E-selectin was assessed by FACS analysis. (C) To demonstrate that the induction of HO-1 by NOD was independent of TNF-α, HUVECs were stimulated for 24 hrs with 10 ng/ml of TNF-α alone, 100 µM of NOD alone or in combination of both. HUVECs cultured in medium served as control. The expression of VCAM-1 and HO-1 was assessed by western blotting. The results shown in A, B and C are representative experiments. A total of 6 independent experiments with different HUVEC cultures were performed. All westernblots have been scanned and statistics was performed on the ratio of the optical density of protein of interest/optical density of GAPDH. Significant inhibition of VCAM-1 expression occurred at concentrations above 12.5 µM and HO-1 induction already at 1 µM (p<0.01).