Differential gene expression between control and athlete subjects after exercise.

<p>(<b>A</b>) Venn diagram representing genes differentially expressed between control and athlete groups at time points T1 (blue; 24 hrs before exercise; n = 4 for each group); T2 (yellow; immediately after exercise; n = 7 for controls and n = 8 for athlete group); and T3 (green; 24 hrs after exercise; n = 4 for each group). Fold change>1.5; False Discovery Rate (FDR)<0.1. Genes differentially regulated at two or three time points are shown in the corresponding intersecting segments. (<b>B</b>) Heatmap representing genes differentially expressed between individual control and athlete samples at time point T2 (immediately after exercise). Each column represents an individual subject. Red represents an up-regulation, and green a down-regulation in gene expression. Probes and samples are clustered by hierarchical clustering Fold change>1.5; FDR<0.1. (<b>C</b>) Top 10 canonical pathway maps representing genes differentially expressed between athlete and control groups at time-point T2 by Metacore analysis with corresponding <i>p</i>-values and FDR.</p

Validation of microarray data.

<p>(<b>A</b>) Fold-change in expression of genes <i>UTS2</i>, <i>HSD11B1</i>, <i>OCLN</i>, <i>IGF1R</i> and <i>INSIG2</i> between athlete (n = 8) and control (n = 7) groups at time point T2, as evident by microarray analysis (Nexus Gene Expression and Genespring), and validated by quantitative real-time PCR (qRT-PCR). qRT-PCR data are normalized to the housekeeping gene and were statistically significant with <i>p</i><0.05 (n = 6 in each group). (<b>B</b>) Concentration of urotensin (pg/ml) in the plasma of control (n = 8) and athlete (n = 6) subjects at time-point T1.</p

¹H NMR spectra of control and athlete urine samples.

<p>¹H NMR spectra of a representative pre- (T1) and post-exercise (T2) urine sample from an athlete (<b>A</b>) and control (<b>B</b>) subject highlighting visible differences between the two spectra and the major resonances. Numbered biomolecules correspond to <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0092031#pone-0092031-t003" target="_blank">Table 3</a>.</p

Physiological characteristics of all participants.

<p>Values are expressed as mean ± SE. “*” indicates statistical significance; p-value<0.01</p

List of differentially expressed genes.

<p>Representative list of genes differentially expressed between control and athlete groups at time point T2. Those indicated in bold have been validated by quantitative real-time PCR.</p

¹H NMR of urine metabolites.

1<p>H NMR identified urine metabolites and relative changes in concentration reported as fold change for athlete and control groups after exercise.</p><p>‡Signal not used for concentration calculation.</p><p>†Peak areas were unreported due to peak overlap.</p><p>♮Metabolite not detected.</p><p>*Significant difference between gr°ups.</p><p>♭Histidine concentrations were not calculated due to dynamic nature of His resonances.</p

Gene expression in control and athlete subjects.

<p>(<b>A</b>) Heatmap of the entire genome including all normalized and filtered probes in athlete and control groups across all time-points (T1: 24 hrs before exercise; T2: immediately after exercise; T3: 24 hrs after exercise). Red represents an up-regulation, and green a downregulation in gene expression. The probes were clustered by k-means clustering into 5 broad clusters. Representative Gene Ontology (GO) biological processes of each cluster are shown in their respective colours on the left. (<b>B</b>) Representative heatmap of probes found significantly different between athlete and control groups across all time-points (T1: 24 hrs before exercise; T2: immediately after exercise; T3: 24 hrs after exercise). The probes were clustered by hierarchical clustering. Fold change>1.5; False Discovery Rate (FDR)<0.1. (<b>C</b>) Top 10 networks representing genes differentially expressed between athlete and control groups across all time points by Metacore analysis, with corresponding <i>p</i>-values and FDR.</p