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Supplemental Information

Figure 1. Body weight of wild-type, CMKLR1 knockout and CMKLR1 heterozygote mice fed on a standard chow diet.

n = 15 for wild-type and heterozygote mice, and 11 and 7 for male and female knockout mice respectively. The upper three lines are for males and the lower three for females.

DOI: 10.7287/peerj.preprints.643v1/supp-1

Suppl. Figure 2. Body fat content of wild-type, CMKLR1 knockout and CMKLR1 heterozygotemice fed on a standard chow diet at 12 weeks (A) and six months (B) of age

n values are as for supplementary Figure 1.

DOI: 10.7287/peerj.preprints.643v1/supp-2

Suppl. Figure 3. Oral glucose tolerance andintraperitoneal insulin tolerance in wild-type, CMKLR1knockout and CMKLR1 heterozygote mice fed on a standard chow diet at 12 weeks (A) and six months (B) of age

n values are as for Supplementary Figure 1. *P<0.05 for knockout and heterozygote compared to wild-type mice of the same sex.

DOI: 10.7287/peerj.preprints.643v1/supp-3

Figure 4. Plasma insulin during the oral glucose tolerance test in wild-type, CMKLR1 knockoutand CMKLR1 heterozygote mice fed on a standard chow diet at 12 weeks (A) and six months (B) of age.

n values are as for Supplementary Figure 1.

DOI: 10.7287/peerj.preprints.643v1/supp-4

Suppl. Figure 5. Insulin tolerance test in6-month-old wild-type, CMKLR1 knockout and CMKLR1 heterozygote mice fed on a standard chowdiet

n values are as for Supplementary Figure 1.

DOI: 10.7287/peerj.preprints.643v1/supp-5

Suppl. Figure 6. Oral glucose tolerance andintraperitoneal insulin tolerance in 6-month-old wild-type, CMKLR1knockout and CMKLR1 heterozygote mice fed on a high fat diet .

n = 17 to 21, except for female knockout mice, where n = 10.

DOI: 10.7287/peerj.preprints.643v1/supp-6

Suppl. Figure 7. Plasma insulin during the oral glucose tolerance test in wild-type, CMKLR1 knockout and CMKLR1 heterozygote mice fed on a high fat diet at 12 weeks (A) and six months (B) ofage.

DOI: 10.7287/peerj.preprints.643v1/supp-7

Suppl. Figure 8. Preliminary experiments on the effects of murine chemerin, its C-15 and C-19 terminal peptides and a stable analogue of its C9-terminal peptide on glucose uptake inmurine mesenteric adipocytes.

Panel (A) shows consolidated results for two preparations of adipocytes which gave very similar basal values and where concentrations of chemerin were identical between experiments; n = 4 to 10. Panels(B) and (C) show means for 4 replicates from single preparations of adipocytes.

DOI: 10.7287/peerj.preprints.643v1/supp-8

Suppl. Figure 9. Body weights (A) and percentage bodyfat (B) of male FVB and C57Bl/6 mice fed on chow and high fat diets for 6 months.

n = 7 to 9. *P< 0.05; **P<0.01; ***P<0.001 for the comparisons indicated by the bars.

DOI: 10.7287/peerj.preprints.643v1/supp-9

Suppl. Figure10. Plasma chemerin concentration (A)and RARRES2 mRNA levels (B) in inguinal adipose tissue of 11- to 12-week-old male db/db mice and the background C57Bl/Ks strain.

n = 6 in A and 5 in B.

DOI: 10.7287/peerj.preprints.643v1/supp-10

Suppl. Figure11. RARRES2 mRNA in perigenital adipose tissue.

(A) Fed and 5 h-fasted C57BL/6 8-week-old wild-type and 10-week-old ob/ob mice, and (B) female ob/ob mice treated with rosiglitazone for 3 weeks. n values and other details are given in the legends to Figures 7D and 8B of the main paper, which, by contrast, show changes in RARRES2 expression in inguinal fat.

DOI: 10.7287/peerj.preprints.643v1/supp-11

Additional Information

Competing Interests

Steven Wang was an employee at AstraZeneca at the time of these studies. AstraZeneca is a pharmaceutical company engaged in the marketing, development and discovery of drugs to treat diabetes.

Author Contributions

Ed T Wargent performed the experiments, analyzed the data, contributed reagents/materials/analysis tools, prepared figures and/or tables.

Mohamed S Zaibi performed the experiments, analyzed the data, contributed reagents/materials/analysis tools, prepared figures and/or tables, reviewed drafts of the paper.

Jacqueline F O'Dowd performed the experiments, analyzed the data, contributed reagents/materials/analysis tools.

Mike A Cawthorne reviewed drafts of the paper.

Steven J Y Wang conceived and designed the experiments, contributed reagents/materials/analysis tools, reviewed drafts of the paper.

Jonathan R S Arch conceived and designed the experiments, analyzed the data, wrote the paper, prepared figures and/or tables, reviewed drafts of the paper.

Claire J Stocker conceived and designed the experiments, performed the experiments, analyzed the data, contributed reagents/materials/analysis tools, wrote the paper, prepared figures and/or tables, reviewed drafts of the paper.

Animal Ethics

The following information was supplied relating to ethical approvals (i.e., approving body and any reference numbers):

1. The University of Buckingham Ethical review Board. 2. Bu08-039; Bu09-007; Bu 11-001; Bu11-009.

Funding

AstraZeneca funded most of the work described in this manuscript (grant 20080509LL/SEML-7DSHX3). Some of the glucose uptake work was funded from the resources of the Clore Laboratory after the formal collaboration with AstraZeneca ended. The original idea to work in this area was Steven Wang's (one of the authors). He had moved from the Clore Laboratory to work for AstraZeneca, who had chemerin receptor knockout mice available to them. The details of the experiments were designed by University of Buckingham authors but discussed with Steven Wang to solicit his input. AstraZeneca played no role in data collection and analysis, the decision to publish, or the preparation of the manuscript, other than it approved the manuscript, asking for no amendments.


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