DGKε knockdown promotes an activated phenotype of ECs. (A) HUVECs and HMECs were transfected with control siRNAs or DGKE siRNAs, and DGKE expression was analyzed after 48 hours at the mRNA level to assess knockdown efficiency. The bar graph shows the mean percentage of inhibition of DGKE mRNA expression (± standard error of the mean [SEM]) from at least 3 independent experiments. **P < .01, ***P < .001. (B) DGKε protein expression was also assessed in cells transfected with control or DGKE siRNAs using western blot analysis. Representative results from 3 independent experiments are shown. (C) ICAM-1 and (D) E-selectin expression at the surface of HUVECs and HMECs was evaluated by flow cytometry 48 hours after DGKE knockdown vs control siRNA–transfected cells. HUVECs stimulated for 4 hours with 100 U/mL TNFα as a positive control for activation. Representative histograms are shown, and the bar graphs illustrate the mean fold change in median fluorescence intensity (MFI) (± SEM) from 4 independent experiments. *P < .05, **P < .01.