Skip to main content
. 2015 Aug 25;5:13524. doi: 10.1038/srep13524

Figure 2. Phosphatase activity of sEH negatively regulates simvastatin-induced NO production and tube formation in ECs.

Figure 2

(a) BAECs were pretreated with the sEH phosphatase inhibitor AFC (50 μM) or ebselen (10 μM) for 2 h or sEH hydrolase inhibitor 12-(3-adamantan-1-yl-ureido)-dodecanoic acid (AUDA, 10 μM), for 2 h and then simvastatin (10 μM) for 24 h. Level of nitrite in culture media was measured by Griess assay. (b–e) BAECs transfected with vector, Flag-tag sEH with PTmut or hydrolase mutant (EHmut); or Flag-tag N-ter or hydrolase domain of sEH (C-ter) for 48 h, then simvastatin (10 μM) for 24 h. (b,c)Western blot analysis of Flag and α-tubulin. (d,e) Level of nitrite in culture media. (f,g) Tube formation with BAECs pretreated with AFC (50 μM) or ebselen (10 μM) for 2 h or transfected with PTmut or N-ter for 48 h, then seeded on pre-coated ECL Cell Attachment Matrix with or without simvastatin (10 μM) for 24 h. Data are mean ± SEM fold branch points from 5 randomly selected microscopy views. *P < 0.05 vs. vehicle- or vector-treated cells, #P < 0.05 vs. simvastatin-treated alone cells.