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Figure 2 RNase protection analysis for the quantification of NOS II- and NOS III-mRNA levels in total RNA from left and right ventricular tissue of NF and failing human hearts from patients with dilated cardiomyopathy (dCMP), ischemic cardiomyopathy (iCMP) or postmyocarditis cardiomyopathy (mCMP). Shown is a representative autoradiogram. Lanes indicate: 1, length marker; 2 to 5, unhybridized antisense-RNA probes; 2, NOS II (619 nt); 3, NOS III (505 nt); 4, Gs (403 nt); 5, all three probes combined; 6 to 23, hybridization of the respective RNA samples with all three antisense-RNA probes; 6, NOS II (587 nt) and NOS III sense-RNA (462 nt); 7, 10 µg total RNA from yeast (negative control), 8 to 22, 20 µg total RNA from NF (8,15,16) and failing human hearts of patients with dCMP (10,12,18,20), iCMP (9,11,14,17) or mCMP (13,19,21,22); 23, 3 µg total RNA from stimulated human hepatocytes (NOS II positive control, 570 nt). Hybridization of total RNA from ventricular tissue with NOS II antisense-RNA, but not with NOS II antisense-RNA, yielded a protected fragment of the expected size (433 nt for NOS III). Sizes of antisense-RNA probes and protected fragments are given in parentheses in the legend.