FIGURE 5.

Levels of pyruvate dehydrogenase complex E1α and E2 proteins, α-ketoglutarate dehydrogenase complex E2 protein, glutamate dehydrogenase protein, and acetyl-CoA acetyltransferases 1 and 2 proteins in human islets are comparable with or higher in human islets than in rat islets or INS-1 832/13 cells. A, levels of PDC E1α protein in human and rat islets are higher than in INS-1832/13 cells. Immunoblot is shown with 15 μg of protein of whole-cell protein/lane probed with anti-PDC E1α antibody and stripped of antibody and reprobed with anti-β-actin antibody to show relative loading of protein across lanes. B, top panel, immunoblot with 20 μg of whole-cell protein/lane. It was probed with affinity-purified primary biliary cirrhosis (PBC) IgG at a 1:20,000 dilution that reacts against the PDC and α-ketoglutarate dehydrogenase complex (KDC) E2 proteins. Lower panel, immunoblot with 5 μg of whole-cell protein/lane probed with the primary biliary cirrhosis IgG at a dilution of 1:30,000 and reprobed with anti-β-actin antibody to indicate the relative total cell protein levels across the lanes. C, two immunoblots. Membranes were probed with anti-glutamate dehydrogenase (GluDH) antibody, stripped of antibody, and reprobed with anti-β-actin antibody to show relative loading of protein across lanes. Lanes in both panels contained 15 μg of whole-cell protein/lane except 3rd lane from left in the upper panel contained 13 μg of protein/lane. D, bottom panels, immunoblots with 15 μg of whole-cell protein/lane (ACAT1, mitochondrial acetyltransferase; ACAT2, cytosolic acetyltransferase).