Fig. 3Figure 4
PDE4D5 expression is upregulated in human failing hearts and PDE4D5 induces hypertrophy and oxidative stress and inhibits mitophagy in NRVMs . A, Representative immunoblots and quantification of PDE4D and PDE4D variants (PDE4D5, PDE4D3, and PDE4D9) expression in left ventricle tissues from healthy donors (n = 3) and heart failure (HF) patients (n = 7). NRVMs were infected with mCherry-PDE4D5 adenovirus (Ad-PDE4D5) or control adenovirus (Ad-Ctrl) for 48 h (B through D ). B, The protein expressions of PDE4D and hypertrophy marker gene, ANP in NRVMs; n = 6 independent experiments. C, Immunofluorescence staining of α-actinin (green) and DAPI (blue) in NRVMs and quantification of myocytes area; n = 24 cells from 3 independent experiments. Scale bar, 20 μm. D, DCFH-DA staining and quantification of ROS levels in NRVMs; n = 5 independent experiments. Scale bar, 200 μm. E, MitoSOX staining and quantification of mitochondrial ROS levels in NRVMs infected with GFP-N-terminal-PDE4D5 a