Fig. 2

Relationship between myocardial ischemia–hypoxia and autophagy-associated proteins in adjacent slices (×400). The positive immunohistochemical staining of a Beclin1, b LC3, c Cathepsin D and d p62 in the cardiomyocytes were stained brown. e The negative control of immunohistochemical staining. The ischemia–hypoxia cardiomyocytes were stained red by f C-2R BG staining. The normal cardiomyocytes stained green by C-2R BG staining f had one-to-one correspondence to p62 positive immunohistochemical cardio myocytes stained brown d. Yellow asterisks indicate the areas of ischemia–hypoxia cardiomyocytes stained red by C-2R BG corresponded to the negative p62 staining (light brown). Red arrows indicate the normal cardiomyocytes, stained green by C-2R BG, corresponded completely to positive p62 staining (brown), and incompletely to positive Beclin1 and Cathepsin D. Positive area of LC3 (d, red arrows) had correlation with nonpositive area of p62 (d, yellow asterisks). As shown in red arrows, there is overlap between positive areas of Beclin1 (a) and Cathepsin D (c)