[Go to this article.]

Figure 3
Primary cardiomyocytes from neonatal rats were stimulated with saline (lane 1) or 30 μM of PE (lanes 2 and 3) in the presence of curcumin (10 μM, lane 3) or a corresponding amount of its vehicle (DMSO, lanes 1 and 2) as a control for 48 hours. (A) Proteins isolated by acid extraction from these cells were subjected to Western blotting for acetylated histone-3/4 or total histone-3/4 as indicated. (B) These cells were subjected to indirect immunofluorescence analysis with antibody against acetylated lysine. Original magnification, ×200. (C) Protein extracts from these cells were subjected to Western blotting with anti-GATA4 antibody, anti-p300 antibody, or anti–β-actin antibody. (D and E) The same extracts (100 μg of protein) were immunoprecipitated with goat anti-GATA4 polyclonal antibody (D) or rabbit anti-p300 polyclonal antibody (E) and subjected to sequential Western blotting with anti–acetylated lysine antibody, anti-p300 antibody, and anti-GATA4 antibody. (F–H) The amounts of GATA4-associated p300/total GATA4 binding (F), acetylated GATA4/total GATA4 (G), and p300-associated GATA4/total p300 binding (H) were quantified by densitometry with the use of Multi Gauge V3.0 (FUJIFILM). The data shown are mean ± SEM from 3 independent experiments.