Adhesion molecules expression analysis.

<p>(A) Relative mRNA levels of VCAM-1, ICAM-1, e-selectin and p-selectin in aortic arches from p55^+/+LDLR^−/− (n = 10) and p55^−/−LDLR^−/− (n = 8) mice. Values are represented relative to expression in p55^+/+LDLR^−/− arches. (B) Immunohistochemical staining of VCAM-1 expression on sections from aortic valve areas indicating a less intense endothelial staining in p55^−/−LDLR^−/− mice. Original magnification×200. (C) Staining quantification. * p = 0.01 by Student's t-test. Error bars indicate SEM.</p

Survival curves, aneurismal incidence and suprarenal aortic diameters.

<p>(A) Percent survival in p55^+/+LDLR^−/− (squares) and p55^−/−LDLR^−/− (triangles) mice following infusion with saline (n = 3 mice/group) or AngII (n = 13–14 mice/group). Comparison of survival curves between AngII-infused p55^+/+LDLR^−/− and p55^−/−LDLR^−/− mice gave a probability value of p = 0.054 by Pearson's chi-square test. (B) representative cross-sections of EVG stained suprarenal aortas from (a) saline infused and of advanced AAA in (b) p55^+/+LDLR^−/− and (c) p55^−/−LDLR^−/− mice indicating dissecting aneurysms and formation of thrombi. Original magnification×100 (a) and×50 (b,c). (C) Percent incidence of AAA in p55^+/+LDLR^−/− and p55^−/−LDLR^−/− mice (p = 0.2 by Fisher's exact test). (D) Suprarenal aortic diameters in p55^+/+LDLR^−/− (n = 12) and p55^−/−LDLR^−/− (n = 8) mice (p = 0.3 by Student's t-test).</p

Atherosclerosis quantification.

<p>(A) Atherosclerotic lesion area in the aortic sinuses of p55^+/+LDLR^−/− (squares, n = 18) and p55^−/−LDLR^−/− (triangles, n = 16) mice. Each symbol represents one animal; bars represent means. * p = 0.02 by Student's t-test. (B) Representative lesions from p55^+/+LDLR^−/− and p55^−/−LDLR^−/− mice are shown. Original magnification×40. (C) Lesion classification according to severity. (D) Gene expression analysis in p55^+/+LDLR^−/− (n = 10) and p55^−/−LDLR^−/− (n = 8) aortic arches. Values are represented relative to expression in p55^+/+LDLR^−/− arches. * p = 0.03 by Student's t-test. Error bars indicate SEM.</p

General characterization of AngII infused mice.

<p>(A) Body weight (B) plasma cholesterol and (C) plasma triglyceride levels in p55^+/+LDLR^−/− and p55^−/−LDLR^−/− mice before (n = 13–14 mice/group) and after 5 weeks of high fat feeding (4 weeks of AngII infusion; n = 8–12 mice/group). (D) Plasma levels of pro-inflammatory cytokines and chemokines (n = 7–9 mice/group) after 5 weeks of high fat feeding (4 weeks of AngII infusion). * p<0.05 by Student's t-test. Error bars indicate SEM.</p

Body weight and plasma lipid levels.

<p>(A) Body weight (B) plasma cholesterol and (C) plasma triglyceride levels in p55^+/+LDLR^−/− (n = 18) and p55^−/−LDLR^−/− (n = 16) mice before and after 8 weeks of high fat feeding.</p

Cytokine and chemokine expression analysis.

<p>(A) Relative mRNA levels of IκBα, TNF, IL-6, IL-10 and (B) MCP-1, MIP-1α, MIP-1β, RANTES in aortic arches from p55^+/+LDLR^−/− (n = 10) and p55^−/−LDLR^−/− (n = 8) mice. Values are represented relative to expression in p55^+/+LDLR^−/− arches. (C) Plasma levels of pro-inflammatory cytokines and chemokines (n = 12–15 mice/group) after 8 weeks of high fat feeding. Error bars indicate SEM.</p

Reperfusion recovery after femoral artery ligation and macrophage treatment (n=9).

<p>A, Representative Laser Doppler images of each group post-operatively and at day 14. B, Treatment with M1, M2a and M2c macrophages increases the reperfusion recovery after 14 days. C, A trend towards an increase in vessel number could be detected in the category ’50-60 µm diameter’ in animals treated with M2a and M2c macrophages.</p

Effect of chlodronate liposomes on blood monocyte and granulocyte population (n=6).

<p>Clodronate liposome treatment reduces the endogenous monocyte population significantly compared to PBS liposome treatment (***: p<0.001) and compared to single clodronate injection (#: p<0.05). A, Single injection with clodronate reduces the monocyte population with 53%, whereas a second injection results in 68% reduction. B, Granulocyte population increases upon clodronate treatment after double injections with clodronate compared to PBS-liposome treatment (***: p<0.001) and single clodronate injection (#: p<0.001), most probably due the depletion of the monocyte population.</p

Reperfusion recovery after clodronate liposome treatment (n=9).

<p>Reperfusion recovery is increased at day 14 in animals treated with either M1, M2a or M2c macrophages compared to NaCl treatment (***: p<0.001). Depletion of the circulating monocytes/macrophages did not inhibit reperfusion recovery and did not alter the effect of exogenously injected polarized macrophages.</p

Effect of inhibiting endogenous macrophage polarization (n=9).

<p>Reperfusion recovery in IL10R1^fl/fl/LysMCre- (WT) and IL10R1^fl/fl/LysMCre+ (KO) mice. Treatment with M2c macrophages improves reperfusion recovery in IL10R1^fl/fl/LysMCre+ mice after 14 days compared to untreated animals (*: p<0.05).</p