Fig. 2

Characterization of adipocyte nuclear corepressor-knockout (ap2-NCoR^flx/flx; AKO) mice. Skin, serum, lung, liver, and adipose tissues were harvested from untreated AKO and littermate control mice on a high-fat diet. Tissues were assessed for histologic and biochemical changes suggestive of alterations in adipocyte function. a Nuclear corepressor (NCoR) expression is significantly reduced in adipose tissue but not in lung, liver, or skin. Tissue was harvested from 3- to 4-week-old mice, and messenger RNA (mRNA) levels were determined by qPCR. The results represent fold changes in triplicate determinations from three or four mice per group and normalized to GAPDH. ** p < 0.01 by Mann-Whitney U test. Black bars, wild-type (WT) mice; white bars, AKO mice. b NCoR IHC staining of perirenal adipose tissue evident in WT mice was not detectible in AKO mice. Arrowheads indicate nuclei with positive staining for NCoR. Representative images are shown. Scale bars = 100 μm. c Quantification of NCoR+ cells in perirenal and intradermal adipose tissue (cells per high-power field [HPF]). The results are derived from 5 HPFs from two or three mice per group. * p < 0.05 by Mann-Whitney U test. d AKO mice have reduced expansion of adipocyte size and increased adipocyte number. Representative images of H&E staining of perirenal adipose tissue from mice fed a high-fat diet. Scale bars = 100 μm. e Quantification of adipocyte size (left) and number (right) in perirenal (top) and intradermal (bottom) adipocytes. Adipocyte diameter and number (area results derived from > 200 adipocytes/mouse and number quantified from 2 to 4 HPF/mouse from three or four mice per treatment group). Data presented are mean ± SD. * p < 0.05 by Mann-Whitney U test. f Ablation of NCoR is associated with increased expression of PPAR-γ-responsive genes. Gonadal adipose tissue was harvested, and mRNA levels were analyzed by qRT-PCR. The results represent fold changes compared with littermate controls. Data were obtained in triplicate determinations from three or four mice per group and normalized to GAPDH. * p < 0.05 by Mann-Whitney U test. g AKO mice have improved insulin sensitivity. The homeostatic model assessment of insulin resistance (HOMA-IR) was calculated from fasting serum insulin and glucose measurements in mice fed high fat diet for 15 weeks, * p < 0.05 by Mann-Whitney U test. h AKO mice have altered circulating adipokine levels. Adipokines were measured in serum of fasting mice using multiplex Luminex assays (n = 4–5 mice/group; * p < 0.05 by Mann-Whitney U test). PEPCK Phosphoenolpyruvate carboxykinase, GLUT4 Glucose transporter type 4, RGS2 Regulator of G-protein signaling 2, PAI-1 Plasminogen activator inhibitor -1