Fig. 2.

CCR5 deficiency reduces M1 gene expression in adipose tissue (AT) during HF diet feeding. Following 16 wk of LF or HF diet, stromal vascular cells were collected from AT and analyzed by flow cytometry. A: representative flow plots of isolated AT stromal vascular cells gated for macrophages and lymphocytes (P1). Cells were further separated by DAPI staining for live and dead cells. Representative flow plot of live cells gated for AT macrophages identified by expression of CD11b⁺F4/80⁺ (n = 3 LF WT; n = 3 LF CCR5^−/−; n = 16 HF WT; n = 14 HF CCR5^−/−) and quantification of AT macrophages. Data in A were analyzed by 2-way ANOVA. B: AT was collected from WT and CCR5^−/− mice. Tissues were stained with an antibody to F4/80 (green) for macrophages and DAPI (blue) for nuclei. Tissues were analyzed by confocal microscopy using a ×40 objective. Sections were chosen from images representing mice with F4/80 gene expression close to the mean of their respective groups. C: AT was collected from WT and CCR5^−/− mice maintained on a HF diet for 16 wk. RNA was isolated and used for real-time RT-PCR analysis, as described in materials and methods. Data are the mean ± SE of the relative gene expression for 12 mice/group. Data in C were analyzed by Student's t-test. *P < 0.05.