Supplemental Data

Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Human , mouse and rat GAL4 reporter assays for PPARγ (A), PPARα (B) and PPARδ (C). % indicates % of maximum response versus appropriate control compound (rosiglitazone for γ, GW7647 for α and GW501516 for δ).
• Supplemental Materials and Methods - Supplemental Materials and Methods for each figure
• Supplemental Figure 2 - The ability of MBX-102 acid to attenuate rosiglitazone induced PPARγ activity was assessed using a reporter gene assay. MBX-102 acid, a weak PPARγ transactivation agonist, was able to inhibit 10 μM rosiglitazone induced PPARγ activation in a dose-dependent manner with an IC₅₀ of 46.7 μM for human PPARγ, 70.3 μM for mouse PPARγ and 56.9 μ M for rat PPARγ
• Supplemental Figure 3 - PPARγ silencing in 3T3-L1 adipocytes. A PPARγ Smartpool siRNA (obtained from Dharmacon) was used to evaluate rosiglitazone and MBX-102 acid stimulated glucose uptake activity in 3T3-L1 adipocytes (as described in the main method section). Rosiglitazone and MBX-102 acid stimulated glucose uptake at a sub-maximal insulin concentration (0.05nM) in 3T3-L1 adipocytes (A) and PPARγハハsilencingハby PPARγ Smartpool siRNA duplexesホハas assessed at the mRNA level (B), fully abolished the enhancement by both PPARγ agonists. Values are plotted as mean ± SEM (n=3, ***, p < 0.001 vs. DMSO-treated adipocytes, 2-way ANOVA and Bonferroni multiple comparison test).
• Supplemental Figure 4 - ZF rat study: Day 43 OGTT results. Plasma glucose (Top panel) and insulin (Bottom panel) were measured at baseline (0 minute), 30, 60, 90 and 120 minutes post-glucose challenge. For clarity, rosiglitazone (left panel) and MBX-102 (right panel) are presented on separate graphs with the ZL and ZF Vehicle Control groups being represented on both graphs. Values represent mean + SEM (n=6-8/group)
• Supplemental Figure 5 - ZF rat study, cumulative body weight gain (A) and cumulative food intake (B).
• Supplemental Figure 6 - 3T3-L1 adipogenesis. MBX-102 acid induced less 3T3-L1 preadipocyte differentiation than rosiglitazone in the presence of insulin. (A): Insulin alone (698nM), (B): Insulin + rosiglitazone (1μM), (C): Insulin + MBX-102 acid (200μM).
• Supplemental Figure 7 - Treatment of murine primary macrophages with MBX-102 acid markedly trans-repressed LPS induced MCP-1, IL-1, IL-1β and IL12p35 cytokine mRNA levels. Values represent mean ± SEM and are representative of at least 2 independent experiments (* = p<0.05, ** = p<0.01, *** = p<0.001, 1-way ANOVA and Tukey's multiple comparison test).
• Supplemental Figure 8 - MBX-102 decreased macrophage infiltration (A), crown-like structures (B) and MCP-1 mRNA levels (C) in epididymal adipose tissue (EWAT) of Zucker Fatty rats.
• Supplemental Table 1 - EC₅₀ (μM) values for MBX-102 acid activation of human, mouse and rat GAL4 reporter assays for PPARγ, PPARα and PPARδ.