Principal features of mouse strains and models.

<p>Principal features of mouse strains and models.</p

Differentially expressed miRNAs by strain.

<p>Dysregulated miRNAs in all mouse models. Positive or negative FC values indicated up or downregulated miRNA respectively compared to control. Listed miRNAs which are “double positive”, i.e. are dysregulated at least 1.5× FC in both AB and Exiqon technology-based screens (except miR-301b in <i>mdx</i> and miR 200a in <i>Mybpc3</i> ki with FC slightly below 1.5). The FC and p values were derived from the Exiqon-based screening. FC values are significant (p≤0.05), except those marked in red.</p

Dysregulated plasma miRNA in DMD patients.

<p>Expression data of 7 miRNAs were studied in a DMD cohort including 5 patients and age-matched controls (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0055281#pone.0055281.s004" target="_blank">Table S3</a>). (A) miRNA expression was normalized to miR-29a and miR-30b, with their respective ROC curves. (B), miRNA expression is presented relative to the miR-31 expression level of each patient, with their respective ROC curves. Individual miRNAs “area under the curve” (AUC) and p values are indicated respectively. (C) DMD plasma creatine kinase and its respective ROC curve. The vertical axe indicates the ratio of the log2 quantities of the tested miRNA relative to the normalizer miRNAs.</p

Hierarchical clustering of 81 serum miRNAs expression level in mouse models for muscular diseases.

<p>The pathological models included in the study are the <i>Sgca</i>-null <i>(Sgca)</i> the <i>Sgcg</i>-null <i>(Sgcg)</i>, the <i>mdx-</i>4CV <i>(mdx)</i>, the wild type C57BL/6 healthy control mice; the KI-<i>Lmna^p.H222P</i> (<i>Lmna</i> ki) and its healthy littermate control (<i>Lmna</i> wt) mice and the KI-<i>Mybpc3^c.G815A</i> (<i>MyBpc3</i> ki) and its healthy control (<i>MyBpc3</i> wt) mice. Total RNA was extracted from serum samples and a two-step miRNA quantification strategy was employed as detailed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0055281#pone-0055281-g001" target="_blank">Figure 1</a>. (A) Valid expression data of 81 miRNAs were subjected to a hierarchical clustering analysis, performed with the Array Studio software. (B) Hierarchical clustering analysis of the three healthy control strains with the expression data of 81 miRNA. (C) Hierarchical clustering analysis of the 5 pathological models strains with the expression data of 22 expressed miRNAs minimally affected by genetic background, listed in (D). Expression level of the miRNAs is presented in color-code with gradient between the dark-blue, representing the lowest expression level, to the deep-read for the highest expression level.</p

Commonly dysregulated miRNAs in all mouse models.

<p>All miRNA species listed are “double positive”, i.e. are dysregulated at least 1.5× FC in both AB and Exiqon technology-based screenings (except miR-200a in KI-<i>Mybpc3^c.A815G</i>, FC = 1.48). FC values are derived from the Exiqon-based screening. The order is of descending FC values in the <i>mdx</i> model. All p values are significant (p≤0.05, Exiqon screen), except for the miR-1 in the KI-<i>Lmna^p.H222P</i> mouse.</p

Serum miRNAs expression and fold change in <i>mdx</i>-4CV and control mouse at the ages of 4 and 22 weeks old.

<p>Expression of 10 miRNAs were studied in the serum of the <i>mdx</i>-4CV mouse at the ages of 4 weeks (mdx4W) and 22 weeks (mdx22W) and in age matched control C57BL/6 mice (B64W and B622W). Serum samples from 6 mice of each strain and time point were pooled and subjected to RT-qPCR quantification using Exiqon technology. Results are the average of three independent experiments. miRNA expression level, designated “abundance” on the vertical axis, is the log₂ of the relative miRNA expression level normalized to miR-29a and 30b, found previously to be the most stable miRNAs in our experimental setup (data not shown). Standard deviation and p values are shown with *stands for p≤0.05, **stands for p≤0.01, and ***stands for p≤0.001. Values in the table are of the corresponding fold change (FC) values for the same miRNAs of the graphical presentation.</p