Is There a Global Reduction of Myofibrillar Protein Density?
For the 2D-gel electrophoresis and immunoblots, equal amounts of protein were loaded onto the gels. This approach would not allow detection of a global reduction of myofibrillar protein density in the myocytes. As myofibrillar proteins make up the majority of total myocyte protein, normalization for total protein concentration of the tissue samples could obscure potential differences. We therefore measured total protein content per volume, as an indicator for the density of myofibrillar protein in the tissue (Figure 5A). Total protein concentration was significantly reduced in HIB compared with CTRL (P<0.05). In another approach we examined the relation between total protein concentration in homogenates and the force developed by the myocytes isolated from the same hearts. This relation showed that the low force developed in HIB corresponded with low protein content (Figure 5B).
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