Figure 1: Generation of transgenic (TG) lines expressing human atrial natriuretic peptide (H-ANP):
A: Western Blot analysis of FLAG-tagged H-ANP (wild-type [WT] and mutant [Mut]) in the TG lines H-WT-NPPA and H-fsMut-NPPA respectively. WT-ANP and Mut-ANP are expressed in all 4 cardiac chambers: right atrium (RA), left atrium (LA), right ventricle (RV), and left ventricle (LV). B: Sequencing of H-fsMut-NPPA-Flag PCR fragments confirms the presence of the genetic variant (T>C), double deletion (delAA) and the extension of the 36 base pairs that results in the anomalous 12-residue carboxyl-terminal tail in Mut-ANP (RITAREDKQGWA). C: Cardiac tissue concentration of Human α-ANP was measured in the purified protein fraction of heart harvested from the 3 groups of mice using ELISA Kit; n=12/group. D: Circulating levels of H-ANP measured in the plasma of the blood collected from the 3 groups of mice using Human-α (1-28 amino acid) ELISA Kit; n=12/group. H-WT-NPPA vs H-fsMut-NPPA; ***P<0001; data is presented as mean ± SD.