Supplementary Figure 1. Transgenic Line 19 displays metabolic syndrome. A) Body weight gain in transgenic line 19 compared with wild type. Every two weeks from Week 4 through Week 18 the body weights of wild type animals (Δ) and transgenic animals from line 19 (▲) were measured. Asterisk indicates p<0.05 for difference between wild type and transgenic body mean body weights. B) Intact wild type (□) or transgenic line 19 (■) mice were weighed, then fat depots from different locations in each were excised and weighed and the weight of fat depots relative to body weight were calculated and plotted as percent of body weight. Epi, epididymal adipose tissue; Mes, mesenteric adipose tissue; Ret, retroperitoneal adipose tissue; SubQ, subcutaneous adipose tissue. Asterisk indicates p<0.05 for difference between wild type and transgenic fat depot weight as a % of body weight. C) Lipoprotein profiles from plasma of wild type (Δ) and transgenic line 19 (▲) mice were determined by size exclusion chromatography. In all cases measurements are mean ± SD from 8 animals.

Supplementary Figure 2. Transgenic line 19 mice have liver steatosis. A) Typical liver from WT mouse (on the left) and from Tg mouse. B)Haematoxylin and eosin stained slices of livers from WT mouse (left) and Tg mouse. C) Oil-red-O staining of liver slices from WT mouse (left) and Tg mouse.

Supplementary Figure 3. Heatmap of gene expression changes in adipose, liver and muscle.Displayed are 3 sets of heat maps derived from 3 tissues: eWAT (epididymal white adipose tissue), liver and muscle. Each set is composed of two subsets of heat maps, one from transgenic line 6 (VP16-1, -2, -3) and one from wild type animals (WT-1, -2, -3). For each tissue / mouse line subset, three heatmaps are displayed, each consisting of a pool of tissue from 3 animals (9 animals total for each line). The Y axis indicates samples and the X axis represents transcripts which were 1.2 fold increased or decreased in tissue from transgenic line 6 relative to the same tissue from wild type animals. The intensity of each pixel is proportional to the relative mRNA levels of one transcript in one sample compared to a baseline, which was formed by pooling WT samples for that tissue. Log ratios between the intensity of each sample and the baseline are shown. Upregulated genes are shown in magenta, downregulated genes are shown in cyan.

Supplementary Table S1.Serum and blood laboratory values of wild type mice and transgenic line 19 mice. Values are expressed as mean + SEM. Hb, haemoglobin; FFA, free fatty acids; TGs, triglycerides; ALT, alanine aminotransferase; AST, aspartate aminotransferase; Ccl2, monocyte chemoattractant protein-1; lymph, lymphocytes; neut, neutrophils; mono, monocytes. n.s.; non-significant. (a) whole blood

Supplementary Table S2. Genes differentially expressed in adipose. Fold change and P value for the comparison Tg vs. WT are shown

Supplementary Table S3. Genes differentially expressed in liver. Fold change and P value for the comparison Tg vs. WT are shown

Supplementary Table S4. Genes differentially expressed in muscle. Fold change and P value for the comparison Tg vs. WT are shown

Supplementary Table S5. Biological processes enriched in genes upregulated in adipose. E value is the P value for enrichment, corrected for multiple testing.

Supplementary Table S6. Biological processes enriched in genes downregulated in adipose. E value is the P value for enrichment, corrected for multiple testing.

Supplementary Table S7. Biological processes enriched in genes upregulated in liver. E value is the P value for enrichment, corrected for multiple testing.

Supplementary Table S8. Biological processes enriched in genes downregulated in liver. E value is the P value for enrichment, corrected for multiple testing.

Supplementary Table S9. Biological processes enriched in genes upregulated in muscle. E value is the P value for enrichment, corrected for multiple testing.

Supplementary Table S10. Biological processes enriched in genes downregulated in muscle. E value is the P value for enrichment, corrected for multiple testing.

Supplementary Methods

11-HSD1 enzyme assay

Activities of human and mouse 11-HSD1 in different fat depots and other tissues were determined by the rate of conversion of cortisone to cortisol. Tissues were homogenized in phosphate buffer containing 5% glycerol and 1 mM EDTA on Qiagen Mixer Mill MM300. Protein concentration was determined using Coomassie Plus Protein Assay (Pierce). Tissue homogenates (50 g/ml protein for liver or 150 ug/ml protein for fat) were incubated with 400 nM 3H-labeled cortisone and 1 mM NADPH in Hepes buffer at 37ºC for 1 hour. The organic phase was extracted with ethyl acetate, dried, resuspended in DMSO, and analyzed on reverse phase HPLC Combiscreen Pro C18 column (Shimadzu). A methanol gradient was used for the elution/separation of cortisone and cortisol, and radioactivity was monitored. HSD-1 activity was expressed as percent conversion and calculated with the formula: %Conversion = cortisol/(cortisol + cortisone), in which the area under the peak of cortisone and cortisol was used. For each homogenate, the activity was determined both in the absence and presence of a mouse HSD-1 specific inhibitor in order to distinguish between the contribution of endogenous mouse HSD-1 and transgenic human HSD-1.