Microarray present call analysis.

<p>A) Average number (Y-axis) and standard deviation of miRNA detected in blood samples of six healthy individuals under four different blood sampling conditions (X-axis). Blood was collected in EDTA blood collection tubes and subsequently transferred into PAXgene^TM tubes at three different time points, i.e. directly (0 min), 10 min, and 2 h after phlebotomy. As control blood was also directly collected in PAXgene^TM blood RNA tubes. B) Venn diagram of miRNAs that were present in all six individuals under different blood sampling conditions. There were 201 miRNAs detected in all individuals under all conditions. No miRNA was detected both in blood that was directly collected in PAXgene^TM blood RNA tubes and in blood that was transferred into PAXgene^TM tubes 2 h after phlebotomy without also being detected in blood directly transferred or transferred 10 min after phlebotomy. C) Balloon plot of miRNAs that show a difference in frequency under the different blood sampling conditions. The X-axis shows the minimal number of individuals that are positive for a miRNA under one condition. The Y-axis shows the maximum number of individuals that are positive for a miRNA under one condition. The top left balloon denotes the 806 miRNAs that were not detected in any of the 24 samples (minimum and maximum of 0). The lower right balloon denotes the 201 miRNAs that were detected in all samples (minimum and maximum of 6). The orange shaded area presents 12 miRNAs that show a difference in frequency of at least three under one of the four conditions as compared to the other conditions. The largest difference was found for miR-769-3p indicated in the lower left corner that was positive in 6 samples under the 2h EDTA condition and not in any sample of the PAXgene condition.</p

Scatter plot of the mean coefficient of variation (CV) for the four blood sampling conditions versus the overall CV.

<p>Both miR-769-3p and miR-135a* showed a higher overall CV as compared to their average CV within each of the four conditions indicating a condition specific abundance of both miRNAs.</p

Box-plots of normalized abundance values of miR-769-3p for the 6 individuals under the four blood sampling conditions.

<p>The most obvious change of abundance of miR-769-3p was found in blood that was transferred in PAXgene^TM blood RNA tubes 2 h after phlebotomy.</p

miRNAs with fold change >1.5 in the different comparisons.

<p>miRNAs in italics are not expressed in all 32 tested samples. Numbers in bold are statistically significant p-values.</p

Comparisons of the fold change of the qRT-PCR and the microarray results.

<p>Comparison of the fold changes of three miRNAs found deregulated in different comparisons in the microarray experiment and the corresponding qRT-PCR results. A fold change means an upregulation of the respective miRNA in the first group of the comparison; a negative fold change means a downregulation of the respective miRNA in the first group of the comparison.</p

Genes deregulated after exercise extracted from Büttner et al. [33] and the miRNAs deregulated in different comparisons in our study that are predicted to regulate these genes.

<p>For certain genes several transcripts are listed. The direction of regulation of the miRNAs and the target genes is given in the last two columns.</p

Anthropometric data and physical capacity.

<p>BMI  =  body mass index; Pmax  =  maximal workload attained during the stepwise incremental exercise test.</p><p>means ± standard deviation; Statistical testing of differences between groups: 1-way ANOVA.</p

Heatmap for the 50 miRNAs with highest variance and all samples analyzed in the study by Radom-Aizik [21] and in our study.

<p>Heatmap for the 50 miRNAs with highest variance and all samples analyzed in the study by Radom-Aizik <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0102183#pone.0102183-RadomAizik1" target="_blank">[21]</a> and in our study.</p

Number of miRNAs expressed in 0–8 samples of each group.

<p>Number of miRNAs expressed in 0–8 samples of each group.</p

Venn diagram of the miRNAs with fold changes >1.5 in the different group comparisons.

<p>Venn diagram of the miRNAs with fold changes >1.5 in the different group comparisons.</p