Figure 2

Immunohistochemistry, immunoblotting and molecular genetic analysis. (A) Micrograph of the muscle biopsy of the right gastrocnemius muscle showing markedly increased variation in fiber size, fiber splitting, increased number of internal nuclei, some very atrophic fibers, and proliferation of adipose and fibrous tissue (hematoxylin and eosin staining, ×100). (B) Dysferlin labeling of the muscle specimen. Arrows point to fibers with diffuse and partly granular cytoplasmic dysferlin accumulation. Arrowheads indicate exemplary fibers with remaining patchy dysferlin signal of the fiber sarcolemma while most fibers completely lack sarcolemmal dysferlin. Bar = 100μM. (C) Immunoblot on muscle lysates from the patient and two controls showing comparable protein expression levels. (D) Sequencing electropherogram of partial anoctamin 5 exon 8 and intron 8 sequences reveals a novel splice-site mutation at intron 8 that may result in skipping of exon 9.