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Supporting information

Supporting Table

Table S1: Conservation of CTRP12 sequences in different vertebrate species. The unknown full-

length sequence of platypus CTRP12 is indicated by ?.

Amino acid identity to mouse CTRP12

Full-length C1q/TNF-like domain

Mouse (Mus musculus) 100 100

Human (Homo sapiens) 70 82

Cow (Bos taurus) 69 78

Horse (Equus caballus) 73 83

Chicken (Gallus gallus) 57 70

Frog (Xenopus laevis) 55 68

Platypus (Ornithorhynchus anatinus) ? 72

Zebra fish (Danio rerio) 52 67

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Fig. S1.Sequence alignment of CTRP12 proteins from different species. Identical residues areshaded in black. Conserved substitutions are shaded in gray.

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Fig. S2. (A-C) Body weights of wild-type C57BL/6 (n=8) (A), ob/ob (n=6) (B), and DIO (n=6)(C) mice overexpressing GFP or CTRP12 over a 10-day period. Adenovirus encoding GFP orCTRP12 was administered on day 0.

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Fig. S3.(A-C) Serum concentrations of TNF- and IL-6 in WT (n=8) (A), ob/ob (n=6)(B), andDIO (n=6) (C) mice overexpressing GFP or CTRP12. (D-F) IL-6 concentrations in WT (n=8)

(D), ob/ob (n=6)(E), and DIO (n=6)(F) mice.

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Fig. S4.CTRP12 does not modulate serum fatty acids, adipose tissue inflammation, or adipocyte size and

number. (A) Serum NEFA levels in WT (n=8), ob/ob(n=6), and DIO (n=6) mice infected with

adenovirus encoding GFP or CTRP12. (B-C) Expression of inflammatory genes in adipose tissue of ob/ob

(n=6) (B) and DIO (n=6) (C) mice expressing GFP or CTRP12. IL-6, Interleukin-6; MCP-1, monocyte

chemotactic protein-1; MIP-1, macrophage inflammatory protein-1; TNF-, tumor necrosis factor-.

(D-E) H&E staining of adipose tissue (200X magnification) derived from the epididymal fat pad of DIO

mice expressing GFP (D) or CTRP12 (E). Representatives of images from adipose tissue sections from

two mice per group were shown. (F) Quantification of cell number using Image J program. Quantificationwas performed by drawing a 2 inch x 2 inch square in the original image file and counting cells within

that square. Ten random fields were chosen in each image.

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Fig. S5. Overexpressing CTRP12 in WT or ob/obmice does not increase Akt (T308) or p44/42-

MAPK (T202/Y204) phosphorylation in skeletal muscle.

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Fig. S6. Examination of the liver function of mice expressing CTRP12. (A) Serum activities of

aspartate aminotransferase (AST) andalanine transaminase (ALT), two common serum markers

of liver damage, were measured in WT (n=8), ob/ob (n=6), and DIO (n=6) mice infected withGFP- or CTRP12-encoding adenovirus. (B) Expression of inflammatory genes (IL-1, IL-6, and

TNF-) in the liver of WT (n=8), ob/ob(n=6), and DIO (n=6) mice expressing GFP- or CTRP12.

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Fig. S7. CTRP12 (10 g/mL) does not induce phosphorylation of IRS-1 (Y612), Akt (T308), orp44/42-MAPK (T202/Y204) in rat L6 myotubes.

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Fig. S8.The expression of CTRP12 in obese prepubertal children compared with lean controlsubjects. Data derived from NCBI GEO database (accession number GDS3688).

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Fig. S9.The specificity of the anti-CTRP12 antibody. Western blot analysis showing that theantibody does not recognize other related CTRPs.