(A) Northern blot analysis with an MCP-1 probe of total RNA isolated from various tissues of 11-week-old MCP-1 Tg-B mice. (B) Plasma concentration of MCP-1 in 13-week-old MCP-1 Tg-B and WT mice. Data are mean ± SEM (n = 7). (C) Immunohistochemical detection of Mac3 in epididymal adipose tissue of 11-week-old MCP-1 Tg-B and WT mice. Macrophages are stained brown. Magnification, ×200 and ×400, as indicated. Scale bars: 50 μm. (D) Macrophage infiltration into epididymal fat tissue was quantitated in MCP-1 Tg-B (n = 7) and WT mice (n = 9) as the ratio of Mac3-positive cells to total cells. Data are mean ± SEM. (E) Quantitation by flow cytometry of the proportion of CD11b+CD45+ cells (macrophages) in the SVF of epididymal fat tissue from 14-week-old MCP-1 Tg-B (n = 7) and WT mice (n = 6). Data are mean ± SEM. (F) Quantitative RT-PCR analysis of total RNA isolated from epididymal fat tissue of 11-week-old MCP-1 Tg-B and WT mice for TNF-α, CD68, and F4/80 mRNAs. Data (mean ± SEM; n = 4) were normalized by the amount of 36B4 mRNA and expressed relative to the corresponding WT value. *P < 0.05, **P < 0.01 versus WT.