Figure 8 .

Autophagy is upregulated in caf mutants but 3-MA treatment does not improve the swimming behavior or dystrophic phenotype of caf mutants. (A) Confocal micrographs of whole-mount embryos stained with anti-LC3 (green), anti-Dystrophin (orange), Phalloidin (magenta) and DAPI (blue). LC3 localization and abundance are affected in caf mutants at 5 dpf; LC3 accumulates at sites of fiber detachment (white arrowheads). (B) Western blot analysis showing increased levels of LC3 II in caf mutants. LC3 II levels were normalized against total protein. (C) Western blot analysis showing decreased levels of LC3 II in WT zebrafish after 3-MA treatment. LC3 II levels were normalized against β-actin. (D–F) Swimming behavior analysis. There was no significant change at 3 dpf in time spent moving (D), total distance traveled (E) or average velocity (F) in caf mutants after treatment with 10 mM 3-MA (n = 165). Bars represent mean ± SEM.