Figure 4.

SERCA2a overexpression increases NOS expression in HCAEC. (a) Reverse trancriptase-PCR analysis of eNOS and SERCA2a mRNAs in HCAEC cells infected with AAV1-S2a or AAV1-βgal for 6 days. β-Actin was used as an internal control. (b) Representative western blot of lysates from noninfected or infected with either AAV1-SERCA2a or AAV1-βgal HCAEC. (c) Histogram showing the relative ratio of eNOS (gray bar), and SERCA2a (black bar) normalized to GAPDH in three experiments. The values were expressed as a percentage of the value obtained for the control in the same blot. The data are mean ± SEM of three experiments (*P < 0.05 versus control). (d) Functional analysis of the human eNOS promoter in HCAEC after infection with Ad-SERCA2a or Ad-βgal. Large-scale analysis of the human eNOS promoter using deletion mutants of 6,047-bp upstream region in fusion with the luciferase gene. Relative promoter activity is expressed as a percentage of luciferase activity in control noninfected cells. Data represent the mean ± SEM of three experiments (*P < 0.05 and **P < 0.01 versus control). AAV, adeno-associated virus; Ad, adenovirus; eNOS, endothelial isoform of nitric oxide synthase; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; HCAEC, human coronary artery endothelial cell; SERCA2a, sarco/endoplasmic reticulum Ca²⁺-ATPase 2a.