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. Author manuscript; available in PMC: 2022 Aug 24.
Published in final edited form as: Circulation. 2021 Jun 23;144(8):615–637. doi: 10.1161/CIRCULATIONAHA.121.053889

Figure 7. Forced sGC activity and NFYA expression rescue exercise induced pulmonary hypertension in CpcPH rats.

Figure 7.

(A) Representative Western blot and quantification of sGCβ1, total NFYA (endogenous 44kDa and DDK-fusion 50kDa), DDK, and GAPDH in PAVSMCs infected with rAAV6-GFP or rAAV6-NFYA-DDK (each n=3). Each plot represents the PAVSMCs sample cultured from one individual rat. (B) cGMP level in PAVSMCs of obese treated with rAAV6-NFYA-DDK or rAAV6-GFP and NO donor DETA NONOate (10μM, 24 hours) (each n=6). (C) Administration of rAAV6-GFP or rAAV6-NFYA-DDK to obese+sugen rats. Representative immunofluorescence images and quantification of NFYA, CD31, αSMA, and DAPI of pulmonary arteries (each n=4). (D) Flow cytometry showing the percent of DDK in CD31 or αSMA positive cells (each n=4). (E) Representative Western blot and quantification of sGCβ1, total NFYA (endogenous 44kDa and DDK-fusion 50kDa), phosphorylation ser 177 eNOS, total eNOS and GAPDH in isolated pulmonary arteries of rats (diameter of PAs: 500–700μm) (each n=4). (F) Quantification of plasma levels of cGMP of the rats (each n=6). (G-H) Right ventricular systolic and end-diastolic pressure (RVSP and RVEDP, each n=6), left ventricular systolic and end-diastolic blood pressure (LVSP and LVEDP, each n=4), and Workload (each n=6) were measured at rest and during exercise (obese+sugen rats infected with rAAV6-GFP or rAAV6-NFYA-DDK). (I-K) Left ventricular Ejection fraction (LVEF), Fraction shortening (LVFS), E wave/A wave ratio (E/A), cardiac index (CI), RVDd, PAAT/ET, TAPSE, and TPRi were measured at rest and during exercise. Rats per group; obese+sugen rats infected with rAAV6-GFP or rAAV6-NFYA-DDK (each n=6). Results are expressed as mean±SEM. *P<0.05, **P<0.01, ***P<0.001. Statistical analyses were performed as described in Figure 1 legend.