Association of GT-repeat length polymorphism with HO-1 expression in atrial tissues.

<p>Representative confocal images show the expression of HO-1 in the atria of 8 AF patients and 4 controls (sinus rhythm subjects [SR], left). The number represents the size of GT-repeats in each subject. Relative intensity of HO-1 measured in the α-actin-expressing areas was quantified (right). S/S denotes 4 of 6 patients with shorter GT-repeat (<27 GT) homozygous genotype and L/L denotes 4 of 6 patients with longer (≧27 GT) homozygous genotype. At least 5 random fields were chosen to observe >30 myocytes with scanning and averaging. Data are expressed as mean ± SE. P<0.05; *, #: the different symbols represent the significant difference among groups.</p

Association of the GT-repeat length polymorphism with fibrosis in AF tissues.

<p>Representative confocal images show fibrosis in the atria of 8 AF patients and 4 controls (sinus rhythm subjects [SR], left). Relative intensity of collagen I was quantified (right). Data are expressed as mean ± SE. P<0.05; *, #: the different symbol represents the significant difference among groups.</p

GT-repeats modulate the transcriptional activity of HO-1 gene and its responsiveness to rapid pacing.

<p>(<b>A</b>) HL-1 cells were transfected with plasmids containing various lengths of GT-repeats in the HO-1 promoter for 24 hours. The luciferase activity was assayed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108773#s2" target="_blank">Methods</a>. (<b>B</b>) HL-1 cells were transfected with plasmids containing various lengths of GT-repeats in the HO-1 promoter for 24 hours and/or subsequently received tachypacing (4 Hz) for 2 hours. Each value (mean ± SE, [n = 4]) is expressed as a fold change of luciferase activity relative to the control condition. P<0.05; *, #: the different symbols represent the significant difference among groups.</p

Effect of PTU on Notch3 and gamma-secretase subunit expression in proximal and distal pulmonary arteries.

<p>Immuohistochemistry shows Notch3 expression in the proximal part (A) (scale bar: 20μm; L: vascular lumen) and the distal part of pulmonary arteries (B) (scale bar: 20μm). Notch3 in medial layer of pulmonary arteries was identified by α-SM-actin double staining for vascular SMC. The picture is a representative of 4 independent experiments.</p

Effect of PTU on pulmonary hemodynamics and pulmonary arterial hypertrophy.

<p>(A) Right ventricle systolic pressure (RVSP) in 4 different groups is shown. Pulmonary hypertension (indicated by elevated RVSP) was established 28 days after MCT injection and PTU treatment (5 mg/100g/day by gavage from day 14 to 28) reduced RVSP in MCT-treated rats. (B) Ratio of RV to LV plus septum weight (RV/LV+S) is shown. (C) Medial wall thickness of small pulmonary arteries (25–50μm) identified by α-SM-actin staining (brown staining) is shown. (D) The degree of medial wall thickness was compared among 4 groups. Each value (mean±SE [n = 6–7]) is expressed. ***p<0.001 versus control or MCT group, one-way ANOVA, bonferroni's post-test.</p

Effect of PTU on gamma-secretase subunit expression in cultured PASMCs.

<p>(A, C) After 24 hours of serum deprivation, PASMCs from control or MCT rats were treated with indicated conditions for 24 hours. Western blot shows gamma-secretase subunit expression in whole cell extraction. (B, D, F) The relative expression level of each protein is quantified by densitometry and normalized to GAPDH. (E) PASMCs from control rats were treated with indicated conditions for 24 hours. Western blot shows gamma-secretase subunit expression in whole cell extraction. Each value represents the mean±SE of 4 independent experiments. *P<0.05, **p<0.01, ***p<0.001 versus control PASMCs without PTU or scrambled peptide or Normaxia, one-way ANOVA, bonferroni’s post test.</p

Demographic and Clinical Characteristics of the Study Population.

<p>ACE = angiotensin converting enzyme; ARB = angiotensin receptor blocker;</p><p>BMI = body mass index; CAD = coronary artery disease; LA = left atrium.</p><p>Demographic and Clinical Characteristics of the Study Population.</p

Association of the GT-repeat length polymorphism with oxidative stress in AF tissues.

<p>An identical paradigm was followed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108773#pone-0108773-g001" target="_blank">Figure 1</a>. Atrial tissues were stained with DHE to detect ROS generation as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0108773#s2" target="_blank">Methods</a>. Representative confocal images show ROS production in the atria of 8 AF patients and 4 controls (sinus rhythm subjects [SR], left). Relative fluorescence density in the α-actin-expressing area was quantified (right). Data are expressed as mean ± SE. P<0.05; *, #: the different symbols represent the significant difference among groups.</p

Characteristics of the experimental study groups.

<p>MCT = monocrotaline; PTU = propylthiouracil; PA = pulmonary artery; T3 = triiodothyronine</p><p>All data are presented as mean ± SE; *p-value<0.05, ^✝p-value<0.01, ǂp-value<0.001versus MCT</p><p>Characteristics of the experimental study groups.</p

Effect of PTU on serum-induced PASMC proliferation and migration.

<p>After 24 hours of serum deprivation, PASMCs from MCT rats were treated with indicated conditions for 24 hours. Translocation of proliferating cell nuclear antigen (PCNA, red staining; scale bar: 75μm) into the nucleus <b>(A)</b> and BrdU incorporation <b>(B)</b> were used to represent proliferative activities of PASMCs. Each value (mean±SE [n = 6]) is expressed as fold change of BrdU incorporation in control cells with 0.1% FBS. (<b>C)</b> Migratory activity of PASMC was assessed by migration assay chamber. After treatment with indicated conditions for 6 hours, PASMCs from MCT rats at the lower aspect of filter membrane was fixed and stained with Liu’s stain (pink staining; magnification 100X). (<b>D)</b> Each value (mean±SE [n = 6]) is determined by cell number at the lower aspect of filter membrane, and expressed as a percentage of control. *p<0.05, **p<0.01, ***p<0.001 versus 10% FBS without PTU, one-way ANOVA, bonferroni’s post test.</p