Altered ADAMTS4 protein levels in cell medium from cardiac cells after stimulation with inflammatory mediators.

<p>Protein levels of ADAMTS4 active forms (p68 and p53) in cell medium after stimulation of IL-1β alone (NFB n = 4, NCM n = 3) and in combination with TNF-α (NFB n = 3, NCM n = 4), relative to controls (n = 4). The bars represent mean levels and the error bars SEM. *p<0.05. NFB, neonatal fibroblasts; NCM, neonatal cardiomyocytes; LPS, lipopolysaccharide; IL, interleukin; TNF, tumor necrosis factor.</p

ADAMTS4 and versican mRNA in cardiac fibroblasts and cardiomyocytes was induced by inflammatory mediators.

<p>mRNA in NFB and NCM of ADAMTS1 (A,B), ADAMTS4 (C,D), and versican (E,F) after stimulation with LPS (n = 6), IL-1β (n = 7), TNF-α (n = 10 (NCM)/6 (NFB)), separate and combined (n = 7 (NCM)/4 (NFB)). Untreated cells served as control (n = 15 (NCM)/18 (NFB)). Box-plots show median (horizontal line), interquartile range (box), 1.5xinterquartile range or maximum/minimum range (whiskers) and outliers (>1.5xinterquartile range). *p<0.05, compared to control. NFB, neonatal fibroblasts; NCM, neonatal cardiomyocytes; LPS, lipopolysaccharide; IL, interleukin; TNF, tumor necrosis factor.</p

Myocardial TIMP-3, HAS and MT4-MMP mRNA levels in rats after aortic banding.

<p>Myocardial mRNA expression of TIMP-3, HAS and MT4-MMP in rats with preserved (HFpFS, n = 9) or reduced FS (HFrFS, n = 6) six weeks after aortic banding, normalized to the reference gene ribosomal protein L4 (RPL4) and relative to sham (n = 9). Box-plots show median (horizontal line), interquartile range (box), 1.5xinterquartile range or maximum/minimum range (whiskers) and outliers (>1.5xinterquartile range). *p<0.05. AB, aortic banding; FS, fractional shortening; TIMP, tissue inhibitor metalloprotease; HAS, hyaluronic acid synthase; MT, membrane-type.</p

Elevated mRNA levels of ADAMTS1, -4 and-8 in rats after aortic banding.

<p>Myocardial mRNA expression of ADAMTS versicanases and aggrecanases in rats with preserved (HFpFS, n = 9) or reduced FS (HFrFS, n = 6) six weeks after AB, normalized to reference gene RPL4 and relative to sham (n = 9). Box-plots show median (horizontal line), interquartile range (box), 1.5xinterquartile range or maximum/minimum range (whiskers) and outliers (>1.5xinterquartile range). *p<0.05. AB, aortic banding; FS, fractional shortening; ribosomal protein L4, RPL4.</p

Versican p150 fragment level was reduced after PPS-treatment in rats with aortic banding.

<p>Myocardial levels of p150 (A) and p70 (B) versican DPEEAE fragment in NaCl-soluble ECM of rats subjected to AB with or without PPS-treatment, shown as representative immunoblot and box-plots with median (horizontal line), interquartile range (box), 1.5xinterquartile range or maximum/minimum range (whiskers) and outliers (>1.5xinterquartile range). *p<0.05. AB, aortic banding; PPS, pentosan polysulfate; ECM, extracellular matrix; HF, heart failure.</p

Elevated mRNA levels of versican and aggrecan in rats after aortic banding.

<p>Myocardial mRNA expression of versican and aggrecan in rats with preserved (HFpFS, n = 9) or reduced FS (HFrFS, n = 6) six weeks after AB, normalized to the reference gene RPL4 and relative to sham (n = 9). Box-plots show median (horizontal line), interquartile range (box), 1.5xinterquartile range or maximum/minimum range (whiskers) and outliers (>1.5xinterquartile range). *p<0.05. AB, aortic banding; FS, fractional shortening; ribosomal protein L4, RPL4.</p

Decreased myocardial ADAMTS4 mRNA level after PPS-treatment.

<p>Myocardial mRNA expression of ADAMTS4 in rats treated with PPS (n = 10) or vehicle (n = 7) six weeks after AB, and in sham-operated rats treated with PPS (n = 3) normalized to the reference gene RPL4 and relative to vehicle-treated sham (n = 3). Box-plots show median (horizontal line), interquartile range (box), 1.5xinterquartile range or maximum/minimum range (whiskers) and outliers (>1.5xinterquartile range). *p<0.05. AB, aortic banding; PPS, pentosan polysulfate; HF, heart failure; RPL4, ribosomal protein L4.</p

Echocardiographic analyses and organ weights in AB-rats.

<p>AB, aortic banding; hypertrophy; HF, heart failure; IVSd, interventricular septal end-diastolic dimension; LVDd, left ventricular diameter at end-diastole; LVDs, left ventricular diameter at end-systole; FS, fractional shortening; PWd, posterior wall dimension; LAD, left atrial diameter.</p

Myocardial mRNA levels after PPS treatment in rats after aortic banding.

<p>Myocardial mRNA expression of ADAMTS1, -5, -8 (A), versican, aggrecan (B), HAS-1, -2, TIMP-3, and MT4-MMP (C) in rats treated with PPS (n = 9 or 10) or vehicle (n = 7 or 9) six weeks after AB, and sham-operated rats treated with PPS (n = 3 or 4) normalized to the reference gene ribosomal protein L4 (RPL4) and relative to vehicle-treated sham (n = 3). The bars represent median levels and the error bars the 75^th percentile. *p<0.05. HF, heart failure, HA, hyaluronic acid.</p

Localization of syndecan-4 to the perinuclear area during muscle cell differentiation.

<p>Myoblasts and myotubes induced to differentiate for three days were fixed with 4% PFA and immunostained with mouse anti-syndecan-4, followed by Alexa 488-conjugated goat anti-mouse (green) before confocal microscopy analysis. Nuclei were stained with DAPI (blue). The framed areas (A-C) are presented at high magnification in the right panels. Arrows denote nuclei with syndecan-4 localized to the nuclear membrane in early differentiated muscle cells (B) and in complex myotubes (C). Scale bar 20 μm.</p