Flow cytometry quantification of EPC following the ISHAGE (International Society for Hematotherapy and Graft Engineering) gating strategy (see Methods section).

<p>Flow cytometry quantification of EPC following the ISHAGE (International Society for Hematotherapy and Graft Engineering) gating strategy (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013790#s2" target="_blank">Methods</a> section).</p

Percentage of circulating CD45^dimCD34⁺KDR⁺ CPC/leukocytes.

<p>(A) in healthy control subjects and patients with stable CAD and ACS. Bars represent mean with SE. (B) in healthy control subjects and patients with stable CAD, troponin T negative (TTneg) ACS and troponin positive (TTpos) ACS. Significance levels on bars represent differences between groups; Significance level for the whole analysis is given above. (C) Correlation of CD45^dimCD34⁺KDR⁺ CPC/leucocytes with the number of diseased (stenosis>50%) coronaries in healthy control subjects and patients with stable CAD and ACS. (D) Correlation of CD45^dimCD34⁺KDR⁺ CPC/leucocytes with the number of individual cardiovascular risk factors in healthy control subjects and patients with stable CAD and ACS.</p

Patients baseline characteristics.

<p>CVRF: cardiovascular risk factor score; Patients baseline characteristics for patients with ACS with troponin positive or negative values are similar.</p

Responses of circulating endothelial progenitor cells (CD45^dimCD34⁺KDR⁺/leukocytes) in patients treated with de novo 40 mg atorvastatin/day.

<p>The left bar indicates the number of progenitor cells before statin therapy, the right bar after 4 weeks of treatment.</p

CX3CR1^-/- leads to enhancement of myocardial fibrosis in Coxsackievirus B3-infected mice.

<p>Bar graphs represent the mean ± SEM of LV (A) TGF-ß1 gene expression, as indicated, after normalization to the housekeeping gene 18S using the 2−^ΔCt formula and normalized to the WT group, which was set as 1, and (B) collagen I/III ratio, demonstrated by immunohistological stainings of (C) collagen I and (D) collagen III. Specific epitopes of collagen I and collagen III are coloured red as shown by representative images (scale bar = 200 μm). Quantification of the positive area (%) / heart area (mm²) was performed via digital image analysis. Statistical analysis was performed by One-way ANOVA or the Kruskal-Wallis test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 with n = 7–12 per group.</p

CX3CR1^-/- increases apoptosis and anti-viral cytokine expression in Coxsackievirus B3-infected mice.

<p>(A) The Bax/Bcl-2 ratio as index for cardiomyocyte apoptosis indicates increased cardiomyocyte apoptosis in infected CX3CR1^-/- mice compared to the corresponding WT mice. (B) Gene expression of anti-viral IFN-β is increased in CX3CR1^-/- CVB3 versus WT CVB3 mice. Bar graphs represent the mean ± SEM. Statistical analysis was performed by One-way ANOVA or the Kruskal-Wallis test. *p<0.05, ***p<0.001, ****p<0.0001 with n = 7–12 per group.</p

CX3CR1^-/- induces left ventricular cell infiltration in Coxsackievirus B3-infected mice.

<p>(A,B) Gene expression level of Ly6C and immunohistological staining of CD68⁺ cells as marker for monocyte infiltration (C,D) Staining for CD3⁺ and CD4⁺ cells in the myocardium after CVB3-infection. The quantitative determination of the immune staining was performed via digital image analysis at a 200x magnification (scale bar = 200 μm). Bar graphs represent the mean ± SEM. Statistical analysis was performed by One-way ANOVA or the Kruskal-Wallis test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 with n = 7–12 per group.</p

CX3CR1^-/- impaired left ventricular function in Coxsackievirus B3-infected mice.

<p>Evaluation of cardiac function via tip catheter. After inoculation with CVB3 CX3CR1^-/- mice show declined heart function as indicated by (A) reduced max. LV pressure (LVP_max), (B) reduced LV contractility (dP/dt_max) and (C) decreased LV relaxation (dP/dt_min). Bar graphs represent the mean ± SEM. Statistical analysis was performed by One-way ANOVA or the Kruskal-Wallis test. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 with n = 7–12 per group.</p

Impact of CX3CR1^-/- on left ventricular chemokine receptor and chemokine expression in Coxsackievirus B3-infected mice.

<p>Bar graphs represent the mean ± SEM of gene expression data of LV (A) CCR2, (B) MCP-1, (C) CX3CR1, and (D) CX3CL1, as indicated, after normalization to the housekeeping gene 18S using the 2−^ΔCt formula and normalized to the WT group, which was set as 1. Quantitative CX3CL1 protein expression (E) was determined via digital image analysis at a 200x magnification (scale bar = 200 μm). Bar graphs represent the mean ± SEM. Statistical analysis was performed by One-way ANOVA or the Kruskal-Wallis test. *p<0.05, ***p<0.001, ****p<0.0001 with n = 7–12 per group.</p

CX3CR1^-/- results in changes in titin isoform phosphorylation.

<p>(A) Expression of the N2B isoform and the corresponding phosphorylation (B). The composition as well as the phosphorylation state of the cardiac titin isoform N2B was determined using Pro-Q Diamond phospho-protein stain. In contrast to N2B phosphorylation the cardiac titin isoform composition is different in CX3CR1^-/- CVB3 mice to WT CVB3 mice, respectively. Bar graphs represent the mean ± SEM. Statistical analysis was performed by One-way ANOVA or the Kruskal-Wallis test. *p<0.05, ***p<0.001 with n = 7–12 per group.</p