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Fig. 6
Inactivation of Washc4 in zebrafish embryos results in skeletal muscle ultrastructural alterations. (a and b) Lateral view of MO1-control- (a) and MO1-washc4-injected (b) embryos showing birefringence at 72 hpf. scale bar: 400 μm. (c) Statistical analysis of birefringence signal. n = 30 (MO1-control), n = 30 (MO1-washc4); two-sided Wilcoxon rank-sum tests. (d and e) H&E-stained sagittal histological sections of skeletal muscle of control embryos (d) and Washc4 morphants (e) at 72 hpf; scale bar = 100 μm. (f to g```) Transmission electron microscopy in MO1-control (f to f``) and MO1-washc4 (g to g```) skeletal muscle at 72 hpf. (f to f``) Control muscle cells exhibit highly organized myofibrils (f``) and normal mitochondria, next to the sarcomeric Z-discs (f`). (g to g``) Washc4 morphant muscle cells show less myofibrils, dysmorphic mitochondria (g``) and an accumulation of various vesicular structures; AV = autophagic vesicle, M = mitochondria, MeB = membranous bodies; scale bar: 2 μm (f, g); scale bar = 1 μm (f` and f``, g` to g```). (j and k) Immunostaining using an anti-acetylated tubulin (acTub) antibody of MO1-control- (j) or MO1-washc4-injected (k) embryos at 72 hpf. scale bar: 200 μm.