Validation of gene expression changes in DHA-treated HUVECs by Western blotting.

<p>(A) HUVECs were treated with DHA for 48 h, after which time total protein were extracted and subjected to Western analysis using an anti-CD47 and an anti-CYP4F2 antibody. As control for equal loading, blots were stripped and re-probed with an anti-β-actin antibody. CD47 and CYP4F2 representative immunoblots are shown in the lower panels, while densitometric analysis for the CD47 and CYPFF2, normalized to β-actin and expressed as fold induction over basal control, are shown in the upper panel. Bars represent mean±SD (n = 3). *P<0.05 vs basal control. (B) HUVECs were treated with/without DHA for 48 h, and then stimulated with IL-1β. After 24 h, total protein were extracted and subjected to Western analysis using antibodies against PDE5α, TGF-β2 and CARD11. As control for equal loading, blots were stripped and re-probed with an anti-β-actin antibody. Representative immunoblots are shown in the lower panels, while densitometric analysis for each protein, normalized to β-actin and expressed as fold over basal control, are shown in the upper panel. Bars represent mean±SD (n = 3). *P<0.05 vs IL-1β.