Representative immunoblots for proteins measured in the right ventricle from normoxic and hypoxic rats.

<p>ACAA2: acetyl-Coenzyme A acyltransferase 2, HADHA: hydroxyacyl-Coenzyme A dehydrogenase/3-ketoacyl-Coenzyme A thiolase/enoyl-Coenzyme A hydratase (trifunctional protein), alpha subunit.</p

Immunoblottings of right ventricle samples from normoxic and hypoxic rats.

<p>Proteins participating in metabolism A: ACAA2, B: HADHA and C: aquaporin 7 show tendency to downregulation by hypoxia. D: monoamine oxidase A, E: tissue transglutaminase and F: endothelin receptor B were all upregulated by hypoxia. ACAA2: acetyl-Coenzyme A acyltransferase 2, HADHA: hydroxyacyl-Coenzyme A dehydrogenase/3-ketoacyl-Coenzyme A thiolase/enoyl-Coenzyme A hydratase (trifunctional protein), alpha subunit. Values are means ± SE and are calculated as percent of normoxia 1 week. n = 6 in both groups at all time points. *P<0.05 vs. normoxia at same time point.</p

Experimental design.

<p>*Three animals from each control and four animals from each hypoxic subgroup were used for GeneChip analysis. Four animals from each group either pulmonary trunk banded (PTB) or sham at week 5 were used for GeneChip analysis.</p

Correlation of the gene chip and qPCR gene expression results.

<p>The correlation is tested by comparing three upregulated, four downregulated and two not regulated genes (according to the gene chip, tsum analysis). There was found significant correlation between qPCR and gene chip analysis. Values are means ± SE. *P<0.05 vs. normoxia, gene chip at same time point, †P<0.05 vs. normoxia, qPCR at same time point.</p

Parameters describing the pulmonary and systemic impact of hypoxia.

<p>SSBP: Systemic systolic blood pressure; HW/BW: Heart weight to body weight ratio.</p><p>n = 6 in both groups at all time points. Values are means ± SE. *P<0.05 vs. normoxia at same time point.</p

qPCR analysis of aquaporin7 and tissue transglutaminase.

<p>A: gene expression of the aquaporin 7 mRNA by use of gene chip and qPCR shows a decrease in gene expression according to hypoxia. B: gene expression of tissue transglutaminase mRNA by use of gene chip and qPCR shows the opposite impact of hypoxia by increasing gene expression. n = 6 in both groups at all time points.</p

Log ratio of gene chip data obtained in the right ventricle of chronic hypoxic compared to normoxic rats versus pulmonary trunk banded (PTB) compared to sham-operated rats.

<p>There was found a positive correlation between regulation of the genes in the two experiments with a significant correlation coefficient R² = 0.69 (P<0.05, n = 288).</p

Right ventricular systolic blood pressure (RVSBP) and right ventricular hypertrophy in hypoxic experiment.

<p>A: changes in RVSBP in the normoxic and hypoxic groups at the four time points. RVSBP was constant in the normoxic group and significantly increased in the hypoxic groups after 2 weeks. B: temporal change in right ventricular hypertrophy in the hypoxic experiment assessed by right ventricular weight relative to body weight ratio (RV/BW) shows significant increase due to hypoxia. n = 6 in both groups at all time points. Values are means ± SE. *P<0.05 vs. normoxia at same time point.</p

Morphometric measurements of the right ventricle.

<p>A: cardiomyocyte diameter was significantly increased by hypoxia compared to normoxic controls. B: examples of staining with reticulin in right ventricle samples from normoxic and hypoxic rats. n = 5–6 in both groups at all time points. Values are means ± SE. *P<0.05 vs. normoxia at same time point.</p

Changes in gene expression by gene chip analysis – hypoxia compared to normoxia.

a<p>Values are the average of a four-way analysis as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0015859#s2" target="_blank">Methods</a>. Only genes with fold changes larger than 2 are included in this table.</p