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Additional Supporting Information may be found in the online version of this article.

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ANA_21926_sm_SupportingFigure1.tif4961KSupplementary Figure 1. Skeletal muscles from hDysf-overexpressing mice show pathological features consistent with myopathy. a, Representative hematoxylin-eosin stained transverse cryosections of proximal and distal muscles from hDysf-high transgenic mice. Scale bar, 100μm. b, Representative hematoxylin-eosin stained section of heart muscle from hDysf-high transgenic mice, revealing areas of calcification. Scale bar, 50μm. Bottom panel is magnification of upper inset panel. c, Representative cross-sections of gastrocnemius muscle from hDysf-high transgenic and wild-type. Scale bar, 100μm. Multiple images of transverse cryosections were examined to quantify fiber cross-sectional area of wild-type and transgenic gastrocnemius muscle (3 animals per genotype, 10 images per animal). The distribution of gastrocnemius fiber size was analyzed between transgenic and littermate wild-type mice, revealing a predominance of smaller fibers in hDysf-mid and –high overexpressing muscle.
ANA_21926_sm_SupportingFigure2.tif3642KSupplementary Figure 2. Morphology and ultrastructure analysis of hDysf transgenic muscle. a, Representative NADH-tetrazolium reductase histochemical staining identifies slow-twitch (dark) and fast-twitch (light) fibers. Fast fibers are selectively affected by hDysf overexpression. b, Electron micrographs of quadriceps muscle from 8-week old wild-type and hDysf-high transgenic mice. Subsarcolemmal vesicles accumulation in hDysf-overexpressing muscle (arrowheads). Left, Scale bar 150nm. Right, Scale bar, 500nm.
ANA_21926_sm_SupportingFigure3.tif5144KSupplementary Figure 3. Sarcolemmal integrity and myoblast fusogenic ability is maintained in hDysf overexpressing mice. a, Evans blue dye uptake in quadriceps and tibialis anterior muscle from wild-type (WT) and hDysf-mid transgenic mice. Scale bar, 200μm. b, Plasma serum levels of creatine kinase in WT, hDysf transgenic and A/J dysferlin-deficient mice (n=5 per group). Values are means ± SD (*P < 0.0001, t-test). c, Fusogenic capacity of wild-type (WT) and hDysf-mid transgenic primary myoblast cultures. Efficiency of fusion was analyzed by quantification of singly-nucleated desmin-positive myoblast cells, cells containing two to three nuclei and myotubes containing four or more nuclei. Five 10X fields from two separate cultures of each genotype were analyzed, comprising 573 hDysf-mid and 512 wild-type nuclei.
ANA_21926_sm_SupportingTable.doc32KSupplementary Table. Primer sequences used for conventional PCR

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