Fig. 1. SR-202 Is a Specific PPARγ Antagonist A, Structure of SR-202 Fig. 1. SR-202 Is a Specific PPARγ Antagonist A, Structure of SR-202. B, Effect of SR-202 on Wy14643-induced PPARα activity. C, Effect of SR-202 on bezafibrate-stimulated PPARβ activity. D, Effect of SR-202 on GW1358-induced FXR activity. E, Effect of SR-202 on troglitazone-induced PPARγ activity. HeLa cells were transfected with the reporter construct (AcoA.TK.CAT or pCAT-promIBABP), and full-length mPPARα, mPPARβ, mPPARγ, or hFXR expression plasmids. They were then treated for 36 h with specific ligands and increasing concentrations of SR-202 as indicated. Each bar represents the mean ± sem of three experiments. CAT values were normalized to the β-galactosidase activity resulting from the expression of a cotransfected vector. From: A New Selective Peroxisome Proliferator-Activated Receptor γ Antagonist with Antiobesity and Antidiabetic Activity Mol Endocrinol. 2002;16(11):2628-2644. doi:10.1210/me.2002-0036 Mol Endocrinol | Copyright © 2002 by The Endocrine Society

Fig. 2. SR-202 Inhibits BRL 49653-Mediated Recruitment of SRC-1 by PPARγ A, Representative gel and quantification by densitometry (CARLA values corresponding to SRC-1 recruitment) of radiolabeled SRC-1 pull down assay, using the hPPARγ ligand binding domain, in the presence of increasing concentrations of BRL 49653. B, Representative gel and quantification (SRC-1 recruitment) of the effect of SR-202 on BRL 49653-stimulated recruitment of SRC-1 by PPARγ. Each bar represents the mean ± sem of at least three experiments. From: A New Selective Peroxisome Proliferator-Activated Receptor γ Antagonist with Antiobesity and Antidiabetic Activity Mol Endocrinol. 2002;16(11):2628-2644. doi:10.1210/me.2002-0036 Mol Endocrinol | Copyright © 2002 by The Endocrine Society

Fig. 3. SR-202 Is a Potent Antagonist of PPARγ-Induced Adipocyte Differentiation A, Effect of SR-202 on BRL 49653-induced adipogenesis of 3T3-L1 cells. Cells were pretreated with SR-202 or vehicle (water) for 24 h and then treated with BRL 49653 (25 nm) and insulin (5 μg/ml), with or without SR-202. After 6 d of treatment, cells were stained with Oil Red O. B, Effect of SR-202 on hormone-induced adipocyte differentiation of 3T3-L1 cells. Cells were pretreated with SR-202 and then induced with dexamethasone/IBMX/Ins with or without SR-202 for 2 d. Thereafter, the medium was changed to insulin alone with or without SR-202 for an additional 2 d. Cells were then stained with Oil Red O. C, Effect of SR-202 on the mRNA levels of aP2 adipocyte differentiation marker. Ribonuclease protection assay was performed on 10 μg of total RNA from 3T3-L1 cells treated as indicated above. L27, Ribosomal protein. From: A New Selective Peroxisome Proliferator-Activated Receptor γ Antagonist with Antiobesity and Antidiabetic Activity Mol Endocrinol. 2002;16(11):2628-2644. doi:10.1210/me.2002-0036 Mol Endocrinol | Copyright © 2002 by The Endocrine Society

Fig. 4. SR-202 Prevents Weight Gain and Adipose Tissue Deposit Content in Mice Body weight (A) and fat weight (B and C) of wt and PPARγ +/− male mice treated or not with SR-202 (400 mg/kg) under both SD and HFD. SR-202 was given as food admixture and the treatment was started just after weaning during a consecutive 10 wk. Each bar represents the mean ± sem of at least 10 mice per group (* and $, P < 0.05; NS, no significative difference). *, Comparison between CTL and SR-202-treated mice; $, comparison between SD and HFD. From: A New Selective Peroxisome Proliferator-Activated Receptor γ Antagonist with Antiobesity and Antidiabetic Activity Mol Endocrinol. 2002;16(11):2628-2644. doi:10.1210/me.2002-0036 Mol Endocrinol | Copyright © 2002 by The Endocrine Society

Fig. 5. SR-202 Treatment Decreases Adipocyte Size A, Histological analysis of epididymal WAT from wt and PPAR γ+/− male mice treated or not with SR-202 (400 mg/kg) under both SD and HFD. Twenty-micrometer sections were stained with both eosin and hematoxylin and with Oil Red O. B, Quantification of adipocyte number (mean ± sem) in at least three microscopic fields of WAT sections per mouse from three different mice per group. *, Comparison between CTL and SR-202-treated mice; $, comparison between SD and HFD (* and $, P < 0.05). From: A New Selective Peroxisome Proliferator-Activated Receptor γ Antagonist with Antiobesity and Antidiabetic Activity Mol Endocrinol. 2002;16(11):2628-2644. doi:10.1210/me.2002-0036 Mol Endocrinol | Copyright © 2002 by The Endocrine Society

Fig. 6. SR-202 Decreases PPARγ Activity in Vivo Amounts of the mRNA of the following PPARγ target genes were determined in WAT of wt and PPARγ +/− mice under HFD treated or not with SR-202: LPL, FAT/CD36, aP2, and SREBP-1c. The mRNA level of each target was determined by reverse transcription followed by real-time PCR using the Light-cycler-FastStart DNA Master SYBR Green I as described in Materials and Methods. Each bar represents the mean ± sem (n = 5). *, P < 0.05 (comparison with CTL wt mice). From: A New Selective Peroxisome Proliferator-Activated Receptor γ Antagonist with Antiobesity and Antidiabetic Activity Mol Endocrinol. 2002;16(11):2628-2644. doi:10.1210/me.2002-0036 Mol Endocrinol | Copyright © 2002 by The Endocrine Society

Fig. 7. SR-202 Decreased Secretion of Adipocytokines Fed levels of plasma leptin (A) and TNFα (B) in wt and PPAR γ+/− male mice treated or not with SR-202 (400 mg/kg) under both SD and HFD. Decreased PPARγ activity, by SR-202 treatment or by deletion of one allele of the PPARγ gene, results in a decrease of leptin and TNFα plasma levels. Each bar represents the mean ± sem of at least 10 mice per group (* and $, P < 0.05; ** or $$, P < 0.01; NS, no significant difference). *, Comparison between untreated and treated mice; $, comparison between SD and HFD. From: A New Selective Peroxisome Proliferator-Activated Receptor γ Antagonist with Antiobesity and Antidiabetic Activity Mol Endocrinol. 2002;16(11):2628-2644. doi:10.1210/me.2002-0036 Mol Endocrinol | Copyright © 2002 by The Endocrine Society

Fig. 8. SR-202 Treatment Protects against HFD-Induced Insulin Resistance Fed levels of plasma glucose (A), insulin (B), and FFA (C) in wt and PPAR γ+/− male mice treated or not with SR-202 (400 mg/kg) under both SD and HFD. Decreased PPARγ activity, by SR-202 treatment or by deletion of one allele of the PPARγ gene, was associated with increased insulin sensitivity. Each bar represents the mean ± sem of at least 10 mice per group (* and $, P < 0.05; $$, 0.01; NS, no significant difference). *, Comparison between untreated and treated mice; $, comparison between SD and HFD. From: A New Selective Peroxisome Proliferator-Activated Receptor γ Antagonist with Antiobesity and Antidiabetic Activity Mol Endocrinol. 2002;16(11):2628-2644. doi:10.1210/me.2002-0036 Mol Endocrinol | Copyright © 2002 by The Endocrine Society

Fig. 9. SR-202 Is an Insulin Sensitizer in Vivo, Decreasing Hyperglycemia and Hyperinsulinaemia in ob/ob Mice Treatment of 8-wk-old ob/ob mice was started at d 0 up to d 20. Serum glucose (A) and insulin (B) levels at the fed state in ob/ob mice on d 0 (white bars) or on d 20 after the treatment with vehicle (gray bars) or SR-202 (black bars). Plasma glucose (C) and insulin (D) levels during an ip glucose tolerance test in ob/ob mice performed at experimental d 20, in untreated mice (white circles) or in mice treated with SR-202 (black circles). E, Intraperitoneal insulin tolerance test in ob/ob mice treated or not with SR-202. F, amounts of the mRNAs of LPL, FAT/CD36, and aP2 in WAT of ob/ob mice treated or not with SR-202 (400 mg/kg) during 20 d. Values are expressed as the mean ± sem (n = 10). *, P < 0.05, **, P < 0.01. From: A New Selective Peroxisome Proliferator-Activated Receptor γ Antagonist with Antiobesity and Antidiabetic Activity Mol Endocrinol. 2002;16(11):2628-2644. doi:10.1210/me.2002-0036 Mol Endocrinol | Copyright © 2002 by The Endocrine Society