Panel A shows cardiac tissue staining with enlarged left and right ventricular cardiomyocyte regions. Panel B shows Western blot images of endothelial nitric oxide synthase and phosphorylated endothelial nitric oxide synthase. Panel C shows a bar graph comparing endothelial nitric oxide synthase expression between left and right ventricular cardiomyocytes. Panel D shows a bar graph comparing phosphorylated endothelial nitric oxide synthase at threonine 495 expression between ventricular regions. Panel E shows a bar graph comparing phosphorylated endothelial nitric oxide synthase at serine 1177 expression between ventricular regions.
Differential phosphorylation of eNOS in LVCM and RVCM. (A) Representative immunohistochemistry showing nNOS staining in transverse sections of adult rat hearts. Low-magnification images indicate the regions of the LV and RV used for higher-magnification views. Enlarged images illustrate representative staining patterns in LVCM and RVCM (scale bar = 2 mm). (B) Representative immunoblots showing total eNOS, phosphorylated eNOS at Thr495 (p-eNOSThr495), and phosphorylated eNOS at Ser1177 (p-eNOSSer1177) in LVCM and RVCM. (C) Quantification of total eNOS expression. eNOS protein levels were significantly lower in RVCM compared with LVCM. (D) Quantification of p-eNOSThr495 expression. No significant difference was observed between LVCM and RVCM. (E) Quantification of p-eNOSSer1177 expression. Phosphorylation at Ser1177 was significantly increased in RVCM relative to LVCM. Each dot represents one biological sample from an individual animal, and one sample per animal per condition was included in the quantitative analysis (N = 16 animals). Source data are available for this figure: SourceData FS1.