Fig 2: Systemic effects of VPC23019 (S1P1/S1P3 antagonist) on obesity, glucose intolerance, and adipocyte hypertrophy/inflammation. A, Body weight change in wild-type (open symbols) and ob/ob (closed symbols) male mice fed the control diet (Con or C) or VPC23019 (40 mg/kg)-formulated diet (VPC or V) after age 10 weeks. B, Daily food intake calculated from 4 cycles of the sum values from 3 consecutive days between weeks 2 and 4 after food change. C, Epididymal fat weights at week 12. D, Intraperitoneal glucose tolerance test (ip-GTT) test in wild-type (left) and ob/ob (right) mice fed Con or VPC diet for 12 weeks. Areas under the curve (AUC) are presented in inset graphs. E to J, Serum levels of E, fasting blood glucose; F, fasting blood (immunoreactive) insulin; G, triacylglycerol; H, total cholesterol; I, aspartate transaminase (AST); and J, alanine transaminase (ALT) in wild-type and ob/ob mice fed the aforementioned diets for 12 weeks. K to N, Representative hematoxylin/eosin (HE)-stained image of K, epididymal and M, inguinal adipocytes and their cell area sizes (L and N, respectively) from wild-type and ob/ob mice. Bars: 100 µm. O, Immunochemical CD11c and CD206 staining of epididymal adipocyte sections prepared from ob/ob mice. The nuclei were stained with DAPI (4',6-diamidino-2-phenylindole). Bars: 50 µm. P, HE-stained hepatic sections prepared from ob/ob mice. Bars: 100 µm. Data are mean ± SD (n in parentheses) and the effects of VPC23019 were not statistically significant in any parameters.

Fig 3: Expression of adipocyte inflammation marker genes in epididymal and inguinal fats from wild-type and ob/ob mice fed the control, JTE-013, SEW-2871, or VPC23019 diet for 12 weeks. Messenger RNA levels of A and B, tumor necrosis factor a (Tnfa); C and D, CD11c (Cd11c); E and F, CD206 (Cd206); and G and H, adiponectin (Adipoq) in the A, C, E, and G, epididymal and B, D, F, and H, inguinal fats of wild-type and ob/ob mice were normalized by that of GAPDH (glyceraldehyde-3-phosphate dehydrogenase) and expressed with that of wild-type mice fed the same control diet as 1. Data are mean ± SD (n in parentheses); *P less than .05 and **P less than .01 vs wild-type samples (black asterisks) or control samples (red and blue asterisks for JTE-013 and SEW-2871 samples, respectively).

Fig 4: Effects of JTE-013 and SEW-2871 on adipocyte hypertrophy and inflammation in epididymal and inguinal fats of wild-type and ob/ob mice. A to D, Representative hematoxylin/eosin (HE)-stained image of cross-sectional A, epididymal and C, inguinal adipocytes and their cell area sizes (B and D, respectively) from wild-type and ob/ob mice fed the control diet (Con), JTE-013 diet (JTE), or SEW-2871 diet (SEW) for 12 weeks. The suppressive effects by JTE-013 or SEW-2871 are statistically significant at *P less than .05, **P less than .01, ***P less than .001 in t test. Bars: 100 µm. E, Immunochemical CD11c and CD206 staining of epididymal adipocytes isolated from ob/ob mice. The nuclei were stained with DAPI (4',6-diamidino-2-phenylindole). Bars: 50 µm. F, HE-stained hepatic sections isolated from ob/ob mice. Bars: 100 µm.