FIG. 3.

Macrophage (MP) infiltration and polarization increase in 3 days of HFD. Mice were treated with NCD (N) or HFD for a series of periods as indicated: 3D, 3 days; 2W, 2 weeks; 10W, 10 weeks. After 10 weeks from when first mice were fed an HFD, mice were killed and epididymal adipose tissues were taken for further analyses. A: Flow cytometry analyses of immune cells in stromal vascular fraction of adipose tissue. #P < 0.05; ##P < 0.01; ###P < 0.001; n = 4 (NCD and 2-week HFD mice) or n = 6 (1-week and 10-week HFD mice). B: Immunohistochemistry analysis of epididymal adipose tissue. The epididymal adipose tissues were stained with antibodies against MAC2 and then subjected to hematoxylin-eosin staining. n = 4. C: Whole-mount immunohistochemistry analysis of epididymal adipose tissue from NCD or 3- or 7-day HFD-treated mice. Samples were stained with DAPI (blue) or antibodies against CD11b (red) and CD11c (green) and visualized by multiphoton confocal microscopy. Scale bar denotes 50 μm. (A high-quality digital representation of this figure is available in the online issue.)