MFAP4-expressing cells.

<p>(A) qPCR analysis of MFAP4 mRNA transcripts in differentiating/contractile VSMCs and cardiomyocytes. MFAP4 mRNA transcripts were highly expressed by differentiating/contractile VSMCs. The data were normalized against β-actin. MFAP4 and β-actin mRNA transcripts were amplified by qPCR with gene-specific primers. (B) SDS-PAGE Western blot analysis using the monoclonal anti-MFAP4 (HG-HYP 7-5) antibody on protein lysate from differentiating/contractile VSMCs and cardiomyocytes. MFAP4 protein was strongly expressed in differentiated/contractile VSMCs as observed by its unreduced state of 66 kDa. Cell culture medium was used as a negative control (NCTR). GAPDH was used as a loading control. (C) SDS-PAGE Western blot analysis using the monoclonal anti-MFAP4 (HG-HYP 7-5) antibody on cell culture supernatant proteins from differentiating/contractile VSMCs and cardiomyocytes. MFAP4 was found in the culture supernatant from differentiated/contractile VSMCs.</p

Spearman correlation analysis of serum MFAP4 and ECM proteins such as fibulin-1, OPG, and OPN.

<p> MFAP4: microfibrillar-associated protein 4; ECM: extracellular matrix; OPG: osteoprotegerin; OPN: osteopontin; Non-STEMI: Non-ST elevation myocardial infarction; CAC: coronary artery calcification.</p

Quantitative real-time PCR analysis of MFAP4 mRNA transcripts in 15 different human tissues.

<p>High expression was detected in the heart, intestine, and lung. Data were normalized against β-actin. Results were calculated as mean+SD values from triplicate measurements. MFAP4 and β-actin mRNA transcripts were amplified by qPCR with gene-specific primers.</p

Immunohistochemical staining of MFAP4 in the basal membrane of the testis (A) and prostate (F) and in the spiral arteries of the uterus (C, E).

<p>Tissues were stained using the monoclonal anti-MFAP4 (HG-HYB 7–14) antibody and counterstained with Mayeŕs hematoxylin. Negative control sections with omission of the monoclonal anti-MFAP4 (HG-HYB 7–14) antibody in the prostate (B) and in the uterus (D). <i>Scale bars:</i> (A, B) 200 µm, (C, D, F) 100 µm and (E) 20 µm.</p

Histological section of a human heart from an area with myocardial infarction and counterstained with Mayeŕs hematoxylin (A).

<p>Immunohistochemical staining using the monoclonal anti-MFAP4 (HG-HYB7–14) antibody and counterstained with Mayeŕs hematoxylin (B). Staining of an infarcted area demonstrating MFAP4 staining within a blood vessel and no staining of cardiomyocytes. <i>Scale bars:</i> (A, B) 100 µm.</p

Immunohistochemical staining of MFAP4 located in elastic fibers in the adrenal gland (A), in the skin (B), in the central arteries and the trabeculae of the spleen (D, E), in blood vessels of the kidney (G), in the liver within blood vessels and the connective tissues in portal areas (H), in the blood vessels located in the submucosal layer and in the lamina propria and of the duodenum (J, K).

<p>Tissues were stained using the monoclonal anti-MFAP4 (HG-HYB 7–14) antibody and counterstained with Mayeŕs hematoxylin. Negative control sections with omission of the monoclonal anti-MFAP4 (HG-HYB 7–14) antibody in the skin (C), in the spleen (F), in the liver (I) and in the duodenum (L). <i>Scale bars:</i> (A, G) 100 µm, (B, C, E, H, I, K) 50 µm and (D, F, J, L) 200 µm.</p

Immunohistochemical staining of MFAP4 in elastic fibers located in the blood vessels and the surrounding connecting blood vessels in the heart (A), in the pulmonary arterioles and interalveolar walls of the lung (C, E) as well as in the lamina propria of the trachea (F).

<p>Tissues were stained using the monoclonal anti-MFAP4 (HG-HYB 7–14) antibody and counterstained with Mayeŕs hematoxylin. Negative control sections with omission of the monoclonal anti-MFAP4 (HG-HYB 7–14) antibody in the heart (B) and in the lung (D) lung. <i>Scale bars:</i> (A, B) 50 µm, (C, D) 100 µm, (E) 20 µm and (F) 200 µm.</p

Demographics of the study population.

<p>¹ Patients with ST elevation myocardial infarction (STEMI).</p><p>² Patients with non-STEMI.</p><p>³ Patients destined for vascular surgery because of various atherosclerotic diseases (stable atherosclerotic disease).</p><p>⁴ Apparently healthy individuals with documented coronary artery calcification (CAC-positive).</p><p>⁵ Apparently healthy individuals without signs of coronary artery calcification (CAC-negative).</p><p>± standard deviation. Mean values </p