Figure 7.

CS exposure enhances adipose tissue macrophage infiltration and MMP12 expression. Subcutaneous adipose was harvested from WT and Mmp12^−/− mice after 6 months of exposure to NS or CS. Adipose tissue from WT mice was analyzed for macrophage infiltration and MMP12 expression (n = 5 mice/group). Representative staining for F4/80+ macrophages (A) and MMP12+ cells (B). The percentages of F4/80+ and MMP12+ cells were calculated relative to methyl green+ and DAPI+ cells, respectively. (C) Representative Western blot for MMP12 in WT mice (80 μg protein/well). Adipose tissue from WT and Mmp12^−/− mice was assessed for endostatin protein in the absence or presence of CS exposure. (D) Representative Western blots for endostatin with densitometric quantification in subcutaneous adipose tissue (E). 3T3-L1 adipocytes were cocultured with subcutaneous adipose tissue homogenates from WT mice exposed to 6-months of NS or CS (n = 3 mice/group). (F) VEGF secretion was quantified by ELISA in the presence or absence of function-blocking antibodies to endostatin or angiostatin and expressed relative to untreated controls. Data are mean ± SEM. *P < 0.05 and **P < 0.005 versus respective NS or untreated controls.