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?(Fig.22). Figure 2. Glut4 mRNA (A) and glut4 protein in the cytosol (B) and membranes (C) of gastrocnemius muscle before and after insulin stimulation in adult rat offspring of dams exposed to ethanol (EtOH) versus control and pair�\fed dams. EtOH + T are EtOH offspring ... Phospho�\Akt and Akt inhibitors Pten and tribbles�\3 in rat muscle Because Akt phosphorylation plays an important role in glut4 translocation to the cell membrane, we determined phospho�\Akt levels before and after insulin stimulation in muscle homogenates (Fig. ?(Fig.3).3). Total Akt and basal phospho�\Akt levels were similar among rat groups. However, this website insulin�\induced Akt phosphorylation at both 473�\serine and 308�\threonine residues was impaired in ethanol rats compared with pair�\fed and control groups, and remained reduced after TUDCA treatment alone. When ethanol rats were treated with insulin in addition to TUDCA, Akt phosphorylation at both 473�\serine and 308�\threonine sites increased to the same level as controls. We next determined PTEN and tribbles�\3 content in muscle homogenates, because these proteins inhibit EX 527 solubility dmso Akt activation by insulin and are increased by ethanol. Both PTEN and tribbles�\3 levels were increased in ethanol rats and were reduced to normal by TUDCA treatment. Figure 3. Akt activation in gastrocnemius muscle in adult rat offspring of dams exposed to ethanol (EtOH) versus control and pair�\fed dams. EtOH + T were EtOH offspring treated with tauroursodeoxycholic acid sodium. (A�CC) Akt and p�\Akt before ... Histone deacetylases in rat muscle We tested whether class I and II HDACs are overexpressed in skeletal muscle of ethanol rats. The mRNA levels of these HDACs were not affected by ethanol or TUDCA treatment (not shown). However, the protein levels of these HDACs showed opposite changes in the nucleus and the cytoplasm. Vatalanib (PTK787) 2HCl We found ~2.5�\fold increased protein content of HDACs of class I (HDAC1 and 3) and class II (HDAC4, 5, 7, and 9) in nuclear extracts of ethanol rats compared with control and pair�\fed groups (Fig. ?(Fig.4).4). TUDCA treatment reduced nuclear HDACs to levels similar to controls. In the cytosol, the HDACs were lower in ethanol rats than in offspring from control and pair�\fed dams, and were increased back to normal levels by TUDCA treatment. Figure 4. Class I and II histone deacetylase (HDAC) proteins in gastrocnemius muscle nucleus and cytosol in adult rat offspring of dams exposed to ethanol (EtOH) versus control and pair�\fed dams. EtOH + T were EtOH offspring treated with tauroursodeoxycholic ... Correlation studies Basal cytosolic glut4 content showed significant (P
