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Fig. 1. Laminin (Lm) and {alpha}-dystroglycan (DG) in normal skeletal muscle. (a-d) Confocal immunofluorescence images of the surface of normal teased muscle fibers immunostained with laminin {alpha}2-G domain (a, FITC secondary, green) and {alpha}-dystroglycan (b, IIH6, Cy3, red) antibodies (a-c, fixed and detergent treated). Laminin and dystroglycan striated patterns colocalize (arrows in a), with less intense laminin fluorescence between striations. (d) Similar fiber image with laminin-dystroglycan costained fiber treated with detergent following laminin immunostaining. (e-h) Peripheral nuclei (e, laminin; f, laminin (green) and {alpha}-actinin (red); g, oblique optical section showing nucleus protruding beyond plane of sarcolemma (dystroglycan, red; {alpha}-actinin, green); h, laminin, green; propidium iodide, red). Peripheral nuclei are sandwiched between {alpha}-actinin of the sarcomere (which coincides with the major costameric circumferential striations) and plasma membrane dystroglycan. (i-j) Periodicity of orthogonally oriented longitudinal optical sections (i, confocal image, laminin-2/4 above and {alpha}-dystroglycan below; j, deconvoluted widefield image, laminin {alpha}2G above and dystroglycan below) of normal fibers. Arrows indicate positions of Z-discs. (k-o) Widefield images reveal repeating pattern of laminin {alpha}2G epitope on internal (k,l) and external (m,n) basement membrane surfaces. (k) An oblique view of an internal basement membrane lying between adjacent fibers contained within a computed three-dimensional pixel (voxel) array prepared from 0.25 µm serial deconvoluted optical sections. (l) Location of internal basement membranes within optical cross-section through three adjacent fibers (arrows). (m-o) Images of fixed teased fiber prepared without detergent and stained with laminin {alpha}2G-specific antibody following (m) and before (n) deconvolution, and incubated only with secondary antibody (o). Costameric elements (Z, M, L striations) and location of peripheral nucleus (n) indicated in m.





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