Figure 3.

SERCA2a gene transfer increases eNOS expression in coronary arteries from MR animals. (a) βgal immunostaining (red) of the left anterior descending (LAD) of a control pig (upper panel) and of a pig injected with AAV1 encoding βgal (lower panel). CD31 (green) was also used in parallel to stain EC. Bar = 50 µm. (b) Representative western blot analysis (n = 3) of SERCA2a and eNOS protein expression in coronary arteries [left circumflex (LCx), right coronary artery (RCA) and LAD] from three experimental groups: sham-operated (gray bar), MR+saline (hatched bar), and MR+AAV1.SERCA2a (black bar). (c–e) Histograms showing relative SERCA2a and eNOS ratios normalized against GAPDH used as a control. The data shown are mean ± SEM (**P < 0.01, *P < 0.05 versus saline); (c) relative expression of SERCA2a in coronary arteries from different group of animals; (d) relative expression of SERCA2a in LAD, RCA, and LCx from different animal groups; (e) relative expression of eNOS in coronary arteries from different animal groups. AAV, adeno-associated virus; βgal, β-galactosidase; EC, endothelial cell; eNOS, endothelial isoform of nitric oxide synthase; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; MR, mitral regurgitation; SERCA2a, sarco/endoplasmic reticulum Ca²⁺-ATPase 2a.