Figure 4

CIDE-A Northern and Immunoblot analyses. A) Northern analysis of RNA extracted from normal (C) and type 2 diabetic (D) liver and heart tissue. Total RNA (10 μg) from the appropriate tissues was resolved by denaturing agarose gel electrophoresis, transferred to positively charged nylon membrane, hybridized with the [α-³²P]dCTP-labeled mouse CIDE-A cDNA and exposed to Bio-Max MR film. Ethidium bromide stain of RNA (10 μg/lane) prior to transfer to nylon membrane. The values represent the level of CIDE-A gene expression for the individual tissue as determined by DNA microarray analysis (ND: not determined). The approximated size (1.3 kb) of the CIDE-A mRNA is noted on the right. B) Immunoblot demonstrating increased CIDE-A protein levels in type 2 diabetic mouse liver. Sixty μg of liver and heart extract was electrophoresed on a 12.5% SDS-polyacrylamide gel and the resolved proteins transferred to a nitrocellulose membrane. The membrane was immunoblotted using a rabbit anti-mouse CIDE-A polyclonal antibody and a goat anti-rabbit IgG polyclonal antibody conjugated to horseradish peroxidase. Arrow indicates mouse CIDE-A. The values represent the level of CIDE-A gene expression for the individual tissue as determined by DNA microarray analysis (ND: not determined).