Ab108816

An ELISA Kit (ab108816, Abcam) was performed to detect the level of BNP, and an ELISA Kit (CEA485Ra, USCN, China) was also performed to examine the concentration of NT-proBNP in mice serum. In addition, the ELISA kits of superoxide dismutases (SOD) (ab65354, Abcam), malondialdehyde (MDA) (ab238537, Abcam), and catalase (CAT) (ab83464, Abcam) were also used according to the manufacturer's protocols.

Morphometry analysis of right ventricular hypertrophy (RVH) was performed by analyzing Fulton’s index (RV/LV+S and RV/BW) and histopathological assessment via Masson’s trichrome staining as previously described.^{22 (link)} Circulating levels of atrial natriuretic factor (ANF) (ab108797, Abcam, Cambridge, UK) and brain natriuretic peptide (BNP) (ab108816, Abcam) were estimated as markers of cardiac hypertrophy using commercially available ELISA kits; the assays were performed according to manufacturer’s instructions. Tissue distribution of ANF was examined as a marker of hypertrophy by immunohistochemistry, and changes in tissue-architecture were analyzed by immunofluorescence staining using α-actin/DAPI. Validation of cHH-induced RVH was performed by analyzing the key molecular pathway for regulation of hypertrophy, that is, the Akt/Gsk-3β-mediated signaling cascade.^{33 (link)}Changes in pulmonary vascular architecture and morphometry post cHH exposure were made in PFA-fixed lung tissue by H&E staining. Tissue sections were microscopically visualized for increments in pulmonary artery medial wall thickness and changes in pulmonary vascular architecture.^{34 (link)}