Figure 3.

Complete deletion of Rnd3 gene resulted in severe apoptotic cardiomyopathy along with the elevation of Rho kinase activity in the mouse hearts. (a) A significant amount of TUNEL-positive cells were displayed in E10.5 Rnd3-null hearts. The arrows indicate TUNEL-positive cells (green) overlapped with nucleus counter-staining. Cardiomyocytes were visualized by red fluorescent staining for F-actin. NS indicates nonspecific staining. (b) Quantification of the TUNEL-positive cells in hearts. (c) Elevated caspase-3 activity was detected in Rnd3-null hearts by immunoblot analyses and quantified by immunoblot densitometry analyses. (d) Elevated Rho kinase activity was detected in Rnd3-null hearts by immunoblotting for two Rho kinase substrates, MYPT1 and MLC2. The immunoblot densitometry analyses were quantified. Statistical significance was determined by unpaired, two-tailed Student's t-test. Data are means±S.D. TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling; MLC2, myosin light chain 2. The numbers in the columns represent the number of mice in each group