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Expression profiling of cardiac genes in human hypertrophic cardiomyopathy: insight into the pathogenesis of phenotypes

^a Section of Cardiology, Department of Medicine, Baylor College of Medicine, Houston, Texas, USA

View larger version (56K): [in a new window]   Figure 1 Subtracted clones dot blotted at 96-spot membranes. Membranes hybridized with radiolabeled forward and reverse subtracted complementary deoxyribonucleic acid libraries. Differentially expressed genes, identified by a twofold increase in signal intensity in the membrane hybridized with the forward subtraction probes, are shown by arrows. Clones hybridized with the reverse subtraction probes are considered background.

View larger version (74K): [in a new window]   Figure 2 Northern blots showing upregulation of expression of a selected differentially expressed gene identified by subtraction hybridization: the upper panels shows blots representing expression of ACTA1, MLC2a, GNAS1, NDUFB10, HSPA8, VDAC1, FHL1 and SARCOSIN in hearts with hypertrophic cardiomyopathy (H) and normal control (N) hearts. The lower panels show the corresponding agarose gels photographs showing 28 S and 18 S ribonucleic acids. ACTA1 = skeletal muscle alpha-actin; FHL1 = four-and-a-half LIM domain protein 1; GNAS1 = GTP-binding protein Gs-alpha subunit; HSPA8 = heat shock 70kD protein 8; MLC2a = myosin light chain 2a; NDUFB10 = reduced nicotinamide adenine dinucleotide ubiquinone oxidoreductase; VDAC1 = voltage-dependent anion channel 1.

View larger version (95K): [in a new window]   Figure 3 Northern blots showing the expression of four differentially expressed genes in the left ventricular tissues of six additional patients with hypertrophic cardiomyopathy (HCM). Lanes 1 and 2 represent the left ventricular samples obtained from normal hearts (donor hearts not used for transplantation). Lanes 3 through 8 represent the left ventricular tissues obtained from patients with HCM. Expression levels of skeletal muscle alpha-actin (A), myosin light chain 2A (B) and GTP-binding protein Gs-alpha subunit (C) were increased in all hearts with HCM, as compared with the control hearts. Expression levels of four-and-a-half LIM domain protein 1 (D) were increased by less than twofold in 4 out of 6 hearts with HCM.