PMC

Metabolic pathways regulated by Acrp30 in the liver. The up-regulation and down-regulation of genes and metabolic pathways are indicated by the respective up and down arrows. Block arrows indicate interactions documented in this report or elsewhere ().

PCR analysis of viral DNA biodistribution in tissues of rats injected PVI with rAAV1-GFP (Upper) or rAAV5-GFP (Lower) vectors. PCR fragments of GFP cDNA (0.6 kb) were visualized by ethidium bromide staining. A negative image of the gel is shown.

RQ RT-PCR assay of the expression levels of mRNAs in the liver of female rats injected PVI with rAAV1-Acrp30 or rAAV5-Acrp30. (A) Expression of PEPCK mRNA. (B) Expression of SREBP-1c. A and B Upper are images of the respective filters with ³²P-labeled RT PCR products, as indicated on the left. A and B Lower are graphic representations of the same data.

Effect of peripherally administered rAAV1-Acrp30 or rAAV5-Acrp30 on glucose tolerance (GT) in DIO female rats. □, DIO control rats injected with rAAV-GFP (n = 10) and fed the HF; ○, control rats injected with rAAV-GFP (n = 6) and fed the ND; ▴, rats injected with rAAV1-Acrp30 vector (n = 6) and fed the HF; ♦, rats injected with rAAV5-Acrp30 vector (n = 6) and fed the HF. IP GTT was conducted at week 28 after treatment (arrows in ). *, P < 0.05, when compared with DIO control rats.

Western blot analysis of proteins in plasma from rats injected with rAAV1-Acrp30 or rAAV5-Acrp30. (A) Plasma from rats at day 40 after treatment. (B) Plasma from rats killed at day 297 after treatment. (C) Same as B, except diluted 1:100 and hybridized to rat-specific anti-Acrp30 antibodies. Plasma from normal mouse was used as a positive control sample (1:100 or 1:200 dilution). Numbers above each lane refer to individual experimental animals. (D) SDS/4–15% PAGE electrophoresis with subsequent immunoblotting. Before loading, prepurified plasma from one of the rAAV5-Acrp30 rats was treated under either reducing (+) or nonreducing (–) conditions. Diluted plasma from mouse, used as a positive control, was treated in the same way. High concentration of proteins in undiluted rat samples under nonreducing conditions resulted in slightly aberrant mobility of the multimer complexes; HMW, high molecular weight; MMW, middle molecular weight; LMW, low molecular weight ().

Effect of peripherally administered rAAV1-Acrp30 or rAAV5-Acrp30 in DIO female rats. All injections, unless indicated otherwise, were done intraportally. (A) Change in BW. (B) Average daily FI shown for all groups. Two groups display significant difference, as indicated. (C) Change in ingested kcal/day. For clarity, only two groups of rats are shown. (D) Average daily caloric intake shown for all groups. (E) Plasma leptin levels at the time that the animals were killed. NS, not statistically significant. (F) Plot of feed efficiency (FE), as explained in Results. □, DIO control rats injected with rAAV-GFP vector (n = 10) and fed the HF; ○, control rats injected with rAAV-GFP vector (n = 6) and fed the ND; •, rats injected intramuscularly with rAAV1-Acrp30 vector (n = 6) and fed the HF; ▴, rats injected with rAAV1-Acrp30 vector (n = 6) and fed the HF; ♦, rats injected with rAAV5-Acrp30 vector (n = 6) and fed the HF. The increase in ingested food and calories documented at the beginning of the experiment (C) is attributed to the switch to the HF, which is more palatable. Arrows in A and C indicate the time when IP GTT was conducted.