Figure 7. The effect of ATF3 overexpression in vitro on hypertrophy and inflammatory markers.

Representative Western blot analysis of cell lysate derived from cultured NRCM transduced with recombinant adenoviruses overexpressing ATF3 or control virus LacZ at the virus amounts of 2, 4, and 8 MOI (A). The expression level of ATF3 (upper panel) and GAPDH loading control (lower panel) from one representative Western blot (performed in triplicates) is shown. RT-qPCR analysis with cDNA derived from mRNA of NRCM cultures transduced with recombinant adenovirus ATF3 or control virus LacZ (B–D, F–G). ATF3 (B), ANP (C), BNP (D), IL-6 (F), PAI-1(G), OSP (H), and BMP-2 (I) mRNA levels are normalized to 18S quantified from the same samples and presented relative to LacZ control cells. The results represent mean ± SEM (n = 27–32) from 3- 4 independent experiments. [³H]leucine (5 µCi/ml) was inserted to culture medium of NRCM transduced with recombinant adenovirus ATF3 or control virus LacZ at the concentration of 4 MOI, and incorporated [³H]leucine was detected by liquid scintillation counter (E). The results represent mean ± SEM (n = 32) from 3 independent experiments. *P<0.05; ***P<0.001.