Journal: Cell metabolism

Article Title: Molecular Integration of Incretin and Glucocorticoid Action Reverses Immunometabolic Dysfunction and Obesity.

doi: 10.1016/j.cmet.2017.08.023

Figure Lengend Snippet: Figure 5. Immunometabolic Effects of GLP-1/Dexa in DIO Mice (A) Selected KEGG pathways enriched in the hypothalamus and Venn diagram showing the overlap of genes significantly regulated by different treatments. Mice (n = 5 per group) were CR or treated with equimolar doses of GLP-1/Dexa, GLP-1, and Dexa for 5 days. The red dashed line represents the statistical significance indicated by the log10 p value (p = 0.05) of a hypergeometric distribution test. ER, endoplasmic reticulum. (B) Representative image showing Iba1- and GFAP-positive cells in the arcuate nucleus of the hypothalamus of mice treated with vehicle or GLP-1/Dexa for 4 days. Scale bar, 100 mm. (C) Quantification of Iba1 (Aif1) and GFAP immunoreactive (ir) cells in the arcuate nucleus of mice treated with vehicle, GLP-1/Dexa, or equimolar doses of GLP-1 and Dexa for 4 days. **p < 0.01 versus vehicle by ANOVA. (D) Analysis of IL-1b levels in the hypothalamus and cortex (n = 8 per group) of mice treated daily with vehicle, GLP-1/Dexa, or equimolar doses of GLP-1 and Dexa for 6 days. *p < 0.05 versus vehicle by ANOVA. Cytokine levels were normalized to the total amount of protein contained in the homogenate. (E) Analysis of plasma cytokines from n = 3 independent experiments by a cytokine antibody array, in mice treated with vehicle, GLP-1/Dexa, or equimolar doses of GLP-1 and Dexa for 6 days. #p < 0.05 comparing compound treatment versus vehicle and *p < 0.05 comparing GLP-1/Dexa versus GLP-1 or Dexa by ANOVA, as indicated in the graph. (C)–(E) are expressed as mean ± SEM.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-iba1 Synaptic Systems Cat. No. 234 003; RRID: AB_10641962 Rabbit polyclonal anti-GFAP DAKO Cat. No. Z0334; RRID: AB_10013382 Biotinylated Goat anti-Rabbit IgG A Vector Cat. No. BA-1000; RRID: AB_2313606 POMC precursor (27-52) (Porcine) Phoenix Pharmaceuticals Cat. No. H-029-30; RRID: AB_2307442 Rabbit anti-Insulin (C27C9) Cell Signaling Cat. No. 3014; RRID: AB_2126503 Guinea pig anti-Glucagon TAKARA Cat. No. M182; RRID: AB_2619627 Goat anti-Somatostatin (D-20) Santa Cruz Cat. No. sc-7819; RRID: AB_2302603 Chemicals, Peptides, and Recombinant Proteins GLP-1 This paper N/A GLP-1/Dexa This paper N/A GLP-1/Dexa (H-C-D) This paper N/A GLP-1/Dexa (C-D) This paper N/A Dexamethasone Sigma-Aldrich Cat. No. D1756 3,30-Diaminobenzidine tetrahydrochloride hydrate Sigma-Aldrich Cat. No. 32750 AraC (cytosine-1-b-D-arabinofuranoside) Sigma-Aldrich Cat. No. C1768 Tamoxifen, Free base Sigma-Aldrich Cat. No. T5648 Paraformaldehyde (PFA) Serva Cat. No. 31628-02 Uranyl acetate Serva Cat. No. 77870 Glutaraldehyde Serva Cat. No. 23114.01 Propylene oxide Sigma Aldrich Cat. No. 82320 Durcupan Sigma Aldrich Cat. No. 44610-1EA Osmium tetroxide EMS Cat. No. 19170 Formvar solution Science Services Cat. No. E15830-25 Lead citrate Science Services Cat. No. E22410 Human Leptin E100 Zhang et al., 1997 N/A Buprenorphine, Buprécare Axience (Pantin, France) Cat. No. 03760087151244 Meloxicam, Metacam Boehringer Ingelheim (Reims, France) Cat. No. 04028691523598 Lidocaine, lurocaine 2% Vétoquinol (Paris, France) Cat. No. 03605877020815 Critical Commercial Assays Vectastain ABC HRP Kit (Peroxidase, Standard) Vector Cat. No. PK-4000 Vectastain Elite ABC HRP Kit Vector Cat. No. PK-6100 C-terminal collagen crosslinks (CTX-1) EIA kit ELISA Immuno Diagnostic Systems Cat. No. AC-06F1 N-terminal propeptide of type 1 collagen (PINP) EIA kit ELISA Immuno Diagnostic Systems Cat. No. AC-33F1 Corticosterone EIA Kit Arbor Assays Cat. No. K014-H1 Mouse Insulin ELISA kit Crystal Chem Cat. No. 90082 Proteome Profiler Mouse Cytokine Array kit Panel A R&D systems Cat. No. ARY006 mTNF HTRF Assay Kit CisBio Cat. No. 6FMTNPEH mIL1b HTRF Assay Kit CisBio Cat. No. 63ADk010pec mIL6 HTRF Assay Kit CisBio Cat. No. 63ADK043PEB Deposited Data RNA sequencing data GEO GEO: GSE102415 Experimental Models: Cell Lines Human: HEK293 cells ATCC Cat. No. CRL-1573 (Continued on next page) e1 Cell Metabolism 26, 1–13.e1–e6, October 3, 2017

Techniques: Clinical Proteomics, Ab Array