Figure 1.

Stimulation of NF-κB activity by various hypertrophic agonists in primary cardiomyocytes. (A) Cardiomyocytes grown on coverslips were serum-starved for 24 h before treatment with PE (50 μM), ET-1 (50 nM), and AngII (50 μM) for 30 min or left untreated. Cells were fixed, blocked, and stained with anti-p65 mAb followed by donkey anti-mouse FITC (The Jackson Laboratory). The nuclear translocation of p65/RelA was visualized by using an Olympus fluorescence microscope. (B) Primary cardiomyocytes were transfected with the 2 × NF-κB reporter plasmid (0.25 μg/plate) and treated with PE (50 μM), AngII (50 μM), and ET-1 (50 nM) for 24 h or left untreated. Cells were harvested, and the Luc activity was determined and normalized to the protein content of each extract. Luc activity expressed by cells transfected with empty vector was given an arbitrary value of 1. The results are presented as the mean ± SE and represent four individual experiments.