Expression of natriuretic peptide receptor genes in C2C12 myocytes and activation of PKG by NPs.

The mRNA levels of (A) myocyte marker genes Myod (B), Pgc1α and NP receptor genes Npra, Nprb and Nprc in differentiating C2C12 myocytes. Data were normalized to internal control gene 36B4. Unpaired two-tailed Student’s t-test, compared with previous day, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001. After differentiation, C2C12 myocytes were serum-starved overnight and treated (D) with membrane-permeable 8-Br-cGMP (200nM) or 8-Br-cAMP (200nM); (E) with ANP (200 nM), BNP (200 nM) or CNP (200 nM) for 30 minutes. Levels of phosphorylated VASP (p-VASP S239), total VASP (VASP), and β-actin were then measured by Western blot analysis.