Figure 4

Downregulation of CARP by GADD153 in the area of IR injury. (A) Photomicrographs of immunohistochemical staining for CARP in the cardiac tissues. Note the significant downregulation of CARP in the rat myocardium after ischemia and reperfusion compared with the sham control (left panels). The immunohistochemical staining for CARP on the endocardium and subendocardial myocardium of the papillary muscle was preserved after IR injury (right panel). Scale bar, 100 µm. (B) GADD153 down-regulates the CARP expression. The cardiomyocytes of neonatal rats were transfected with 2 µg of the expression vectors for GFP-GADD153 or GFP. The cells were fixed with 4% paraformaldehyde and permeabilized in 0.4% Triton X-100 for 10 min at room temperature. The cells were stained for 15 min with 0.1 µg/ml Hoechst33258 to visualize the nucleus (blue) and for indirect immunofluorescent staining against CARP (red). The cells were then stored in 50% glycerol in PBS at 4℃. The GFP fluorescence (green) was analyzed using confocal microscopy. Trans, transmission. Scale Bar, 20 µm.