Figure 4. The effects of deletion of the AIE and the C-terminal tail on eNOS phosphorylation.

The cell lysates from various eNOS–HEK-293 stable clones, including empty pCDNA3.1(+), WT eNOS (WT), Δ594–604 (Δ594), Δ605–612 (Δ605) and Δ1164–1177 (Δ14) were immunoblotted with antibodies specific to phosphosites at (A) p-Thr⁴⁹⁵, (B) p-Ser⁶¹⁵, (C) p-Ser⁶³³ and (D) p-Ser¹¹⁷⁷. (E) An antibody specific to total eNOS was used to confirm equal expression levels for the various eNOS constructs. The blots shown are representative of at least three experiments. Densitometry was used to quantify the phosphorylated eNOS relative to total eNOS of respective constructs. Data are presented as the percentage of WTeNOS and represent means±S.D. (*P<0.05; **P<0.01; NS, no significant difference versus WT eNOS).