Fig. 7.

In vitro assessment of left ventricular endothelial nitric oxide synthase (eNOS) and NO bioavailability in Normal and DM + HFD + CKD swine. Left ventricular endothelial eNOS mRNA (a) and protein (b) levels, as well as phosphorylation (p-eNOS) at the Ser1177 site (c) were not different between groups, while p-eNOS at the inhibitory Thr495 site was lower in DM + HFD + CKD swine (d). No differences between groups in eNOS glutathionylation (e), coupling (f), myocardial nitric oxide metabolites nitrite (NO₂⁻) and nitrate (NO₃⁻, g) and the ratio of phosphorylated to non-phosphorylated vasodilator activated phosphoprotein (p-VASP/VASP, h) were observed. Values are mean ± SEM. The original western blots are available in the online supplemental file: original eNOS and p-eNOS (Ser1177) western blots are shown in Supplementary Fig. 6, p-eNOS (Thr495), VASP and p-VASP blots are shown in Supplementary Fig. 7, Glutathione/eNOS blots are shown in Supplementary Fig. 8, and eNOS monomer/dimer blots are shown in Supplementary Fig. 9. *P < 0.05 for DM + HFD + CKD versus Normal by Student’s t test