Figure 2
Rnd3+/− haploinsufficient mice were predisposed to hemodynamic stress. Severe cardiac apoptosis and Rho kinase activation were detected after TAC. (a) Significant increases in TUNEL-positive cells were observed in Rnd3+/− hearts compared to WT control hearts after TAC. The arrows indicate TUNEL-positive cells (green) overlapped with nucleus counter-staining (blue). Cardiomyocytes were visualized by red fluorescent staining for F-actin. Scale bar represents 10 μm. (b) Quantification of the TUNEL-positive cells in mouse hearts. Statistical significance was determined by one-way ANOVA followed by Student–Newman–Keuls method. Data are means±S.D. (c) Higher caspase-3 activities were detected in Rnd3+/− hearts compared to WT animal hearts after TAC. (d) Densitometry analysis of active caspase-3 normalized by GAPDH. (e) Hyperphosphorylation of Rho kinase substrate MYPT1 exhibited in Rnd3+/− hearts indicates elevated Rho kinase activity compared to WT hearts after TAC. (f) Densitometry analysis of MYPT1 phosphorylation. Statistical significance was determined by unpaired, two-tailed Student’s t-test. Data are means±S.D. Casp3, caspase-3; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; MYPT1, myosin phosphatase target subunit; ROCK1, Rho-associated coiled-coil kinase 1; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling. The numbers in the columns represent the number of mice in each group
