Overexpression of mini-agrin in skeletal muscle increases muscle integrity and regenerative capacity in laminin-2-deficient mice

Supplemental Data

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• Figure S1 - Transcript levels of β-actin and muscle creatine kinase (MCK) are not changed in the different mouse models. A) Relative expression of β-actin using mRNA encoding MCK to normalize the amount of RNA used. Transcripts encoding β-actin show a trend to be slightly increased in dy^3K/mag-tg and dy^3K/dy^3K mice, which may reflect the relative increase of non-muscle cells in the diseased muscle. However, this difference was not statistically significant. B) Relative expression of mRNA encoding MCK using β-actin for normalization. Again, a nonsignificant trend towards a decrease in MCK mRNA was observed in dy^3K/mag-tg and dy^3K/dy^3K mice. Data represent the mean ± SEM (N=6). Based on these results, all the measurements for laminin-α5, -α4 and dystroglycan were normalized to the average value obtained for β-actin and MCK (see Material and Methods). (pdf, 36 KB)
• Figure S2 - Tenascin-C immunoreactivity in tibialis anterior in 4-week-old mice. In dy^3K/dy^3K mice, many regions positive for tenascin-C were seen while only few such regions were detected in dy^3K/mag-tg. No tenascin-C immunoreactivity was detected in wt mice. Bar = 50 µm. (pdf, 30 KB)
• Figure S3 - H & E staining of tibialis anterior 5 days after notexin injection. In dy^3K/dy^3K mice, the majority of the muscle contained mononucleated cells (asterisk) and only a few, very small muscle fibers (arrowhead) were found. In contrast, regenerating muscle in dy^3K/mag-tg and wt mice contained larger regenerating muscle fibers (arrowheads) and much fewer mononucleated cells. Bar = 20 µm. (pdf, 125 KB)