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. 2018 Oct 23;7(21):e009624. doi: 10.1161/JAHA.118.009624

Figure 9.

Figure 9

Effects of sapropterin (tetrahydrobiopterin) on superoxide production and endothelial NO synthase (eNOS) dimer/monomer protein levels in embryonic day (E) 12.5 hearts. A, Representative images of dihydroethidium staining in the ventricular myocardium of E12.5 hearts from control and diabetic dams with and without tetrahydrobiopterin administration. A subset of heart sections from diabetic dams were pretreated with Nω‐nitro‐l‐arginine methyl ester (L‐NAME; 300 μmol/L) or superoxide dismutase (SOD; 100 U/mL) for 30 minutes before dihydroethidium probing. B, Quantification of dihydroethidium fluorescence signals. C, Representative Western blotting showing eNOS dimer and monomer bands of E12.5 hearts from control and diabetic dams with and without tetrahydrobiopterin treatment. Denatured endothelial cell lysate validates the size of the eNOS monomer band. D, Densitometric analysis of eNOS dimer/monomer ratios. n=5 to 6 hearts per group from 2 to 4 litters. Data are means±SEM and analyzed using 2‐way ANOVA, followed by Bonferroni post hoc test. EC indicates endothelial cell. *P<0.05 vs untreated control; † P<0.05, †† P<0.01 vs untreated diabetes mellitus.