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Figure S1. Effects of taurine on plasma levels of triglyceride (TG) and cholesterol in obese mice. Fasting plasma levels of TG (A) and cholesterol (B) at the end of treatment period. Results are presented as the mean ± SEM (n = 6). Statistical comparisons were made with each vehicle control; *P < 0.05 was considered significant.

Figure S2. Taurine suppressed hyperglycemia development in obese diabetic KK-Ay mice. Four-week-old male diabetic obese KK-Ay/Ta Jcl mice were purchased from CLEA Japan (Tokyo, Japan). All the animals were kept in individual cages in a temperature-controlled room at 24°C ± 1°C under a 12 h light/dark cycle. After 1 week of adaptation period, The KK-Ay mice were fed a HFD containing 60% fat or a HFD supplemented with taurine (2% or 5% w/w) for 4 weeks. The energy intake of all the mice was adjusted by pair feeding. Adipose tissues and blood samples were obtained at the end of the treatment period. (A) Body weight change. Body weight was measured every week in KK-Ay mice fed a HFD or HFD containing taurine. Each data point represents the mean ± SEM (n = 6). (B) Fasting plasma glucose in the KK-Ay mice fed HFD with or without taurine for 4 weeks. Results are presented as the mean ± SEM (n = 6). Statistical comparisons were made with each vehicle control; *P < 0.05 was considered significant. (C-E) The mRNA expression levels of F4/80, TNF-α and Arg1 in epididymal adipose tissues of control (0%) and taurine-treated mice were quantified by real-time PCR. Results are presented as the mean ± SEM (n = 6). Statistical comparisons were made with each vehicle control; *P < 0.05 was considered significant.

Figure S3. Effects of taurine on IL-4 induced alternative M2 activation in BMDM. (A) Representative flow cytometry results of CD11c and CD206 marker expression in BMDM treated with PBS as the vehicle control, IL-4 (10 ng/ml), or IL-4 plus 400 μM taurine for 20 h. (B) The mRNA expression levels of M2 macrophage markers in BMDM treated with PBS as the vehicle control or 10 ng/ml IL-4 for 20 h were quantified by real-time PCR. The relative amount was normalized to the amount of the 36B4 transcript. The value of a vehicle control was set at 100%, and the relative value was presented as fold induction to that of the vehicle control. Results are presented as the mean ± SEM (n = 4). Statistical comparisons were made with each vehicle control; *P < 0.05, **P < 0.01 was considered significant.

Figure S4. Expression of myeloperoxidase (MPO) in adipose tissue. (A) Increase in adipose tissue MPO expression in response to a HFD. Male C57BL/6J mice at 6 weeks of age were fed a HFD. At 0, 3, 7, 14 and 21 days, mice were sacrificed and epididymal adipose tissues were dissected out, and then lysed by lysis buffer containing 20 mM Tris-HCl (pH 7.5), 15 mM NaCl, 1% Triton X-100 and a protease inhibitor cocktail (Nacalai Tesque). The protein concentration was determined using the DC protein assay reagents (BioRad Laboratories), and 80 μg of protein was separated by 10% SDS-PAGE and transferred to a polyvinylidene difluoride (PVDF) transfer membrane (GE Healthcare UK). After blocking, the membrane was incubated overnight at 4°C with anti-MPO (Santa Cruz Biotechnology, CA, USA), and then with a secondary antibody conjugated to horseradish peroxidase (anti-goat IgG [Promega, WC, USA]) for 1 h. The secondary antibody staining was visualized with a chemiluminescence immunoassay using ImmunoStar LD (WAKO chemicals, Osaka, Japan). Results shown are representative of three independent experiments. (B) Expression of MPO in adipose tissues of taurine-fed mice. Epididymal adipose tissues of C57BL/6J mice treated without taurine (0%) or with taurine (2% or 5%) for 14 weeks while on a HFD were dissected out and lysed. The expression of MPO in 3 individual mice of each group was analyzed as described above. Results shown are representative of two independent experiments.

Table S1. Oligonucleotide sequences for primers used for RNA analysis.

Table S2. Plasma taurine level and taurine concentration in tissues of taurine-fed mice. Taurine concentration in plasma, liver, and epididymal, mesenteric and subcutaneous white adipose tissue (WAT) of C57BL/6J mice treated without taurine (HF; control) or with taurine (HF+Tau 2% or 5%) for 14 weeks while on a HFD. The values are means ± SEM of 3–4 samples. *P < 0.05, compared with each HFD control (0%).

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