Fig. 1.
Endothelial nitric oxide synthase (eNOS) RNA and protein expression is decreased by ∼90% in the late pregnant (LP) NG-nitro-l-arginine methyl ester (l-NAME)-treated (LPLN) main uterine artery (mUA). A: LP and LPLN mUA were dissected, placed immediately in Trizol reagent for subsequent RNA extraction, and quantitative (q)PCR analysis as described in materials and methods. Hprt1 and Ywhaz were used as control genes for the qPCR. eNOS RNA expression was significantly decreased in the LPLN mUA compared with LP. B: LP and LPLN mUA were dissected and placed in modified RIPA buffer for protein analysis as described in materials and methods. Twenty-five micrograms of protein were loaded on the gel and GAPDH was used as a loading control. Top: representative immunoblot. Bottom: densitometry was used to quantify the immunoblots probed for eNOS and normalized by GAPDH expression. eNOS is expressed relative to GADPH. eNOS is significantly decreased in LPLN-treated mUA. *P < 0.05 by Student's t-test; n = 3 animals per group.