(A) Relative protein expression levels of p-eNOSS1177 and eNOS after 50nM sPRR-His 2-h and 3-h treatment were measured by Western blotting. (B) Effect of 50nM sPRR-His on relative protein expression levels of p-eNOSS1177 and eNOS in the presence or absence of 10μM Los. (C) Effect of 50nM sPRR-His on NO generation in the presence or absence of 10μM Los. (D) Effect of 50nM sPRR-His on relative protein expression levels of p-eNOSS1177 and eNOS in the presence or absence of AT1R siRNA knockdown. (E) Effect of 50nM sPRR-His on NO generation in the presence or absence of AT1R siRNA knockdown. The value beneath the image indicates the densitometry of the protein normalized to eNOS for 3 separate experiments. N=9 (N represents the number of samples in each group, and the repetitions of separate experiments are 3). Statistical analysis was performed by using one-way ANOVA with the Bonferroni test for multiple comparisons or by using unpaired Student’s t test for two comparisons. *P<0.01, compared with the CTR or Scr siRNA; #P<0.01, compared with the sPRR-His or sPRR-His+Scr siRNA; ##P<0.05, compared with the sPRR-His or sPRR-His +Scr siRNA. DAF (diaminofluorescein)-FM indicates 4-amino-5-methylamino-2′, 7′-difluorofluorescein diacetate. Data are mean ± SEM.