Fig. 3

Measuring TNF-α protein levels inside the HUVECs using immunofluorescence staining (a). Data revealed that Pal induced the synthesis of TNF-α in HUVECs 48 h, indicated by green-colored appearance. The addition of Insulin and DHA decreased the TNF-α levels showing a reduced inflammatory response. Detection of NO and nitrosative status using Griess assay (b; n = 3). According to our data, Pal induced nitrosative stress in HUVECs by the increase of NO. The addition of DHA and insulin reduced NO production in these cells and returned to near-to-control levels. One-Way ANOVA and Tukey posthoc analysis (n = 3). *p < 0.05; **p < 0.01; ***p < 0.001; and p < 0.0001