8/11/2019 jbc.M111.303651-1
1/10
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
8/11/2019 jbc.M111.303651-1
2/10
Fig. S1.Sequence alignment of CTRP12 proteins from different species. Identical residues areshaded in black. Conserved substitutions are shaded in gray.
8/11/2019 jbc.M111.303651-1
3/10
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.
8/11/2019 jbc.M111.303651-1
4/10
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.
8/11/2019 jbc.M111.303651-1
5/10
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.
8/11/2019 jbc.M111.303651-1
6/10
Fig. S5. Overexpressing CTRP12 in WT or ob/obmice does not increase Akt (T308) or p44/42-
MAPK (T202/Y204) phosphorylation in skeletal muscle.
8/11/2019 jbc.M111.303651-1
7/10
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.
8/11/2019 jbc.M111.303651-1
8/10
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.
8/11/2019 jbc.M111.303651-1
9/10
Fig. S8.The expression of CTRP12 in obese prepubertal children compared with lean controlsubjects. Data derived from NCBI GEO database (accession number GDS3688).
8/11/2019 jbc.M111.303651-1
10/10
Fig. S9.The specificity of the anti-CTRP12 antibody. Western blot analysis showing that theantibody does not recognize other related CTRPs.
Top Related