Figure 2. Neutrophils recruited in the femoral artery of wt mice by HFD.

(a) Time schedule of the neutrophil depletion experiment. Wt mice were treated with the neutrophil depletion antibody 1A8 or the control IgG at 22 days and 24 days after starting HFD. Leukocyte recruitment was observed in these mice 28 days after HFD (left); 1A8 clearly diminished neutrophils in peripheral blood compared with IgG (right). (b) Leukocyte recruitment in the femoral artery of wt mice fed HFD treated with 1A8 (right image) or IgG (left image). Bar, 50 μm. The graph shows that leukocyte recruitment was significantly inhibited by neutrophil depletion (n = 4 and 4, respectively). Data are presented as the mean ± SEM. ****P < 0.0001 by unpaired; 2-tailed Student’s t test. (c) Monocyte depletion in the blood of LysM-eGFP mice using clodronate liposomes. The highest expression of GFP in peripheral leukocytes was observed in neutrophils, but monocytes were intermediately positive as well. Monocytes were then depleted using clodronate liposomes. Treatment of clodronate liposome-depleted GFP⁺Ly-6C^hi leukocytes as monocytes (right) in peripheral blood, but not GFP⁺Ly6G⁺ leukocytes as neutrophils (left). (d) Leukocyte recruitment in HFD-fed LysM-eGFP mice treated with clodronate liposomes to deplete monocytes. Clodronate liposomes were administered 24 h before observation. The image on the left shows the femoral artery treated with vehicle and that on the right shows the femoral artery treated with clodronate liposomes. Bar, 50 μm. Leukocyte recruitment did not change with monocyte depletion (graph) (n = 4 and 4, respectively). Data are presented as the mean ± SEM. NS (not significant) by unpaired; 2-tailed Student’s t test.