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Figure 4 Western blots of CHAPS-solubilized protein from nonfailing (NF) and failing human heart tissues from patients with dilated cardiomyopathy (dCMP), ischemic cardiomyopathy (iCMP) and postmyocarditis cardiomyopathy (mCMP). Electrophoreses were performed in 7.5% resolving gels and blots were immunostained with anti-NOS II antibody (upper panel) or anti-NOS III antibody (lower panel). For normalization of the NOS staining signal, blots were simultaneously immunostained with an antibody to beta-tubulin. Positive control proteins were included (CHAPS-solubilized homogenates, from lipopolysaccharide-induced RAW 264.7 macrophages for NOS II and from EA.hy 926 endothelial cells for NOS III). Protein samples were applied as follows: heart muscle samples (75 µg protein/lane), RAW 264.7 macrophages and EA.hy 926 endothelial cells (20 and 40 µg protein/lane, respectively). Results are representative of four independent experiments with identical results.