Figure 4.

Cytosolic and nuclear compartmentalization of Akt in diaphragm skeletal muscle with mechanical ventilation A, cellular fractionation of proteins. Whole skeletal muscle homogenates were fractionated into cytosolic and nuclear proteins for Western blot analysis. Representative Western blot of nuclear histone proteins for verification of cellular fractionation. Cytosolic (Cyt) and nuclear (Nuc) protein fractions were subjected to electrophoresis, transferred to nitrocellulose membranes and subsequently probed for expression of the nuclear compartmentalized histone protein 1 (H2B) or the cytosolic compartmentalized copper zinc superoxide dismutase (CuZnSOD) protein. B, representative Western blot and ponceau- stained nitrocellulose membranes to demonstrate even loading and transfer for the analysis of Akt in the cytosolic and nuclear compartments of skeletal muscle. C, percentage total cytosolic phosphorylated (% phospho) Akt protein. D, percentage total nuclear phosphorylated (% phospho) Akt protein. Values are percentages of total phosphorylated protein abundance and presented as means ± s.e.m.* Significantly (P < 0.05) different from control values. † Significantly (P < 0.05) different from 6 h MV values. ‡ Significantly (P < 0.05) different from 6 h MVT values.