Blockade of monocyte binding to endothelial cells by APC in different concentrations under flow conditions.

<p>Monocyte binding to HUVECs in dynamic adhesion assays after 5 min (left) and 10 min venous flow (middle), and after 1 min arterial flow (right). Pre-treatment of HUVECs with 10 µg/mL activated protein C (drotrecogin alfa) (dark grey bars) diminished monocyte adhesion compared to 1 µg/mL (light grey bars) and 5 µg/mL (grey bars) drotrecogin alfa. (^##<i>p</i><0.005; ^#<i>p</i><0.05 vs. 1 µg/mL APC).</p

Monocytes bind sEPCR in a concentration dependent manner.

<p>Monocytes bind soluble, recombinant EPCR in a concentration dependent manner in flow cytometry analysis. GST tagged-EPCR was detected with an Alexa Flour-labeled anti-GST antibody. Secondary antibody and isotype IgG served as controls (black bars on the left and second from left). (*** <i>p</i><0.0001; # <i>p</i><0.05 vs. controls).</p

Equal blocking of monocyte binding to endothelial cells by anti-EPCR and APC under flow conditions.

<p>Monocyte binding to HUVECs in dynamic adhesion assay after 5 min (left) and 10 min venous flow (middle), and after 1 min arterial flow (right). Pre-treatment of HUVECs with anti-EPCR (dark grey bars), anti-Mac-1 (light grey bars), or with activated protein C (drotrecogin alfa; crosshatched bars) diminished monocyte adhesion to an equal extent compared to control. (*** <i>p</i><0.0001; ^###<i>p</i><0.0005; ^##<i>p</i><0.005; ^#<i>p</i><0.05 vs. no blocking; ns  =  statistically not signifiant).</p

Specific binding of CHO cells expressing activated Mac-1 to recombinant EPCR in static adhesion assay.

<p>Specific binding of CHO cells transfected with permanently activated Mac-1 (Mac-1+ CHO cells; black bars) to soluble, recombinant EPCR in a static adhesion assay (left). Blocking with an anti-Mac-1 antibody resulted in loss of EPCR binding capacity of Mac-1+ CHO cells (right). Native CHO cells without transfection of Mac-1 served as a negative control (light grey bars). (^###<i>p</i><0.0005; ns  =  statistically not significant vs.native CHO cells; *** <i>p</i><0.0001).</p

TRAF6 deficiency does not alter lipid deposition in the abdominal aorta.

<p>Lethally irradiated 6 week old TRAF6^+/+/LDLR^−/− mice received TRAF6-deficient (hatched bars, N = 10) or competent fetal liver cells (white bars, N = 10), TRAF6^+/−/LDLR^−/− mice received TRAF6-deficient fetal liver cells (black bars, N = 10) only. Subsequently, all groups consumed high cholesterol diet (HCD) for 18 weeks. Abdominal aortas were fixed in formalin, pinned, and stained with oil red O to detect lipid deposition. Oil red O-positive staining in per cent of total area is shown as mean±SEM in the upper panel (A), representative images are shown below (B).</p

TRAF6 deficiency does not modulate atherogenesis in mice.

<p>Lethally irradiated 6 week old TRAF6^+/+/LDLR^−/− mice received TRAF6-deficient (hatched bars, N = 21) or competent fetal liver cells (white bars, N = 21), TRAF6^+/−/LDLR^−/− mice received TRAF6-deficient fetal liver cells (black bars, N = 22) only. Subsequently, all groups consumed high cholesterol diet (HCD) for 18 weeks. Intimal lesion area of the atherosclerotic plaques in aortic roots was quantified. Pooled mean intimal lesion area ± SEM are shown as graphs in the upper panel (A), representative sections stained with oil red O below (B).</p

TRAF6 expression in blood does not associate with acute or chronic coronary heart disease.

<p>325 patients undergoing coronary angiography were divided into three groups: no coronary heart disease (No CHD, N = 77), stable coronary heart disease (CHD, N = 178), and acute coronary syndromes (ACS, N = 70). TRAF6 and GAPDH mRNA was analyzed by quantitative real-time PCR in total blood RNA. Results are presented as mean±SD computed from the average measurements obtained from each group.</p

Subtypes of blood leukocytes.

<p>Subtypes of blood leukocytes.</p

TRAF6 deficiency does not modulate inflammatory reactivity of macrophages toward cholesterol and palmitic acid.

<p>Bone marrow-derived macrophages were isolated from 6 week old TRAF6^+/+/LDLR^−/− mice receiving TRAF6-deficient (hatched bars, N = 4) or competent fetal liver cells (white bars, N = 4), were stimulated with 4 mg/ml cholesterol or 0.75 µM palmitic acid, and assayed for expression of IL-6 (A), MCP-1 (B), TNFα (C), and IL12-p70 (D) by cytometric bead array.</p

Demographic and clinical characteristics of study participants.

<p>*, p<0.05 vs controls.</p><p>∫, p<0.05 vs CHD.</p