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Exogenous NAD+ improves the structure of muscle in dag1 morphants. (A-B) Anterior left, dorsal top, side-mounted 2 dpf embryos stained for actin (phalloidin, red) and laminin-111 (green or white). Qualitatively, laminin-111 antibody staining appears to be within myotomes and less well aligned at the MTJ BM in dag1 morphants (A) compared to dag1 morphants treated with 0.1 mM NAD+ (B). White boxes in (A) and (B) correspond to numbered panels below. White arrowheads indicate holes in the MTJ BM. (A3, B3) 2DWTMM analysis of laminin-111 stained dag1 morphants (A3) and NAD+-supplemented dag1 morphants (B3). Maxima nodes are in red, maxima chains are in blue, and vectors pointing in the direction of the maximum intensity gradient are in green. Parallel vectors reflect greater organization. (C) Quantification of the anisotropy factor. The anisotropy factor is the sum of the vector angles. A greater anisotropy factor denotes more organization. NAD+ treatment of dag1 morphants (blue bars) significantly increases organization of laminin-111 compared to dag1 morphants (gray bars) over multiple size scales; *p<0.05, **p<0.01. (D) Model of the MTJ. Transmembrane receptors, integrins, and the DGC bind extracellular laminin. In dag1 morphants, laminin is less organized at the MTJ BM. Exogenous NAD+ improves laminin organization in the MTJ BM in Dag1-deficient zebrafish. Scale bar is 50 micrometers.
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