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Fig. 5. Reduced fibrosis in the Mstn^-/- mice. (A) Low-power micrograph (i), and high-power micrograph (ii) of hematoxylin and eosin staining (H&E) and Van Geisen (iii) staining of day 28 (D28) wild-type and Mstn^-/- muscle sections; bar, 100 µm. Bar, 10 µm. Thick connective tissue (arrows) is only seen in wild-type muscle sections with H&E staining (ii). Similarly, considerably more collagen (red) staining was seen in the wild-type muscle sections (iii; Van Geisen), indicating over-expression of extracellular membrane proteins, leading to fibrosis. Further evidence of increased fibrosis was observed in the scanning electron micrographs of wild-type muscle after 24 days of regeneration (iv); bar, 120 µm. (B) Increased expression of decorin mRNA in the Mstn^-/- regenerated muscle. (i) Quantitative RT-PCR was performed on total RNA isolated from day 28 wild-type and Mstn^-/- muscle and the amplicons were detected by southern hybridization. (ii) Densitometry analysis and normalization of results to GAPDH expression indicates that there is increased expression of decorin mRNA in the regenerating Mstn^-/- muscle.

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