Additional file 2: Figure S2. of Identification in GRMD dog muscle of critical miRNAs involved in pathophysiology and effects associated with MuStem cell transplantation

Clinical follow-up. Clinical scores of mock GRMD dogs (---) and MuStem cell-injected dogs (—) are represented as mean ± SD. The clinical score of each GRMD dog was assessed weekly and expressed as a percentage of a theoretical healthy dog score. Limits of the MuStem cell delivery window are indicated (dashed lines). (PDF 32 kb

Clinical follow-up.

<p>(A) Mean±SD clinical scores of mock GRMD dogs and MuStem cell-injected dogs (GRMD^MuStem). The clinical score of each GRMD dog was assessed weekly and expressed as a percentage of a theoretical healthy dog score. Limits of the MuStem cell delivery window are indicated (dashed lines). (B) Right lateral view of a GRMD^MuStem dog, #7G^Mu. (C) Right lateral view of mock GRMD dog, #4G. Note the anterior weight transfer and plantigrady.</p

Flow chart of the statistical filtration of microarray data.

<p>Procedure leading to classification for each of the three pairwise comparisons of healthy, GRMD and GRMD^MuStem dog muscle samples.</p

Description of the ten dogs used in the <i>in vivo</i> study.

<p>Description of the ten dogs used in the <i>in vivo</i> study.</p

Histological analysis of the <i>Biceps femoris</i> muscle of 9-month old dogs.

<p>Healthy (#3H), GRMD (#6G), and GRMD^MuStem (#7G^Mu) dog muscles are presented respectively in left, mid and right panel. (A, B, C) Muscle tissue presentation after hemalun eosin safran staining. (B) Mock GRMD dogs display a typical dystrophinopathic pattern with diffuse anisocytosis, hypertrophic hyaline fibres (open arrowhead), centronucleated fibres (arrow), necrotic fibres (*), regenerative foci (black arrowhead) and multifocal thickening of the endomysial space by fibrosis (highlighted by the saffron yellow staining). (C) The dystrophinopathic pattern is remodelled in GRMD^MuStem dogs: anisocytosis is milder, hypertrophic hyaline fibres and necrotic fibres are less numerous, whereas centronucleated fibres are more abundant (arrow) compared to GRMD dogs. Inset, another picture taken on the same tissue sample with a higher magnification that displays centronucleated fibre. (D, E, F) Regenerative activity of muscle fibres in healthy, GRMD and GRMD^MuStem dogs, as indicated after immunolabelling specific to the developmental isoform of the myosin heavy chain (MyHCd). (G, H, I) Dystrophin expression in healthy, GRMD and GRMD^MuStem dogs. (A-B-C; G-H-I) Scale bar = 200 μm (in set Scale bar = 100 μm). (D, E, F) Scale bar = 500 μm.</p

Functional characterization of GRMD dog model.

<p>A selection of Gene Ontology (GO) terms significantly enriched in the groups of genes up- and down-regulated between GRMD dog model and healthy dog. Associated with the GO term, the total numbers of genes assigned with this annotation are indicated, followed in the rectangle by the number of genes observed (on left) and expected by chance (on right). Numbers in bold indicate a significant over-representation of the corresponding GO term. Enrichments are colour-coded according to the associated p-value from red (enriched) to blue (depleted).</p

Impact of MuStem cell systemic delivery on GRMD dog muscle.

<p>(A) False-colour heatmap illustrating the transcripts significantly differentially expressed between GRMD^MuStem dog and mock GRMD dog samples. Each line and column corresponds to a probe and an individual biological sample, respectively. The gene names assigned to each probe are indicated on the right, separated by a comma in the case of multiple names as for example with the myosin isoforms. The number of probes / genes are indicated beside each expression profile name. The standardized log₂-transformed intensities are plotted with a colour scale ranging from -2.50 (blue) to 2.50 (red) incremented by 0.25. (B) RT-qPCR validation of 15 genes. For each gene, fold change values are indicated for GRMD^MuStem dog <i>versus</i> GRMD dog. (C) Histograms of RT-qPCR for two selected genes: MUSTN1 and GPD1. Relative quantifications (RQ) are calculated by normalizing with respect to RPS18.</p

Comparison with human studies.

<p>Comparison with human studies.</p