Fig. 6

Skeleton length is less disrupted in dag1 morphants than in fkrp morphants and is not significantly improved with NAD+ supplementation. (A–C6) Anterior left, dorsal top, side-mounted embryos at 72 hpf with labeled actin, AChRs, and SV2. (Lettered panels) Phalloidin stained embryos. (1) Merged channels of AChR and SV2. (2) Skeletonized images. (3) Magnification of phalloidin channel. (4) Magnification of SV2 channel. (5) Magnification of AChR channel. (6) Magnification of skeleton channel. (A–A6) Control embryo. (B–B6) dag1 morphant embryo. (C–C6) dag1 morphant embryo treated with NAD+ at 6 hpf. (D) Length of skeletons per myotome in control embryos (n = 186 half-myotomes), dag1 morphants (n = 260 half-myotomes), and dag1 morphants receiving NAD+ (n = 252 half-myotomes) at 6 hpf. Skeleton length is not significantly different between untreated and NAD+ treated dag1 morphants. (E) Degree of branching within the myotome in control embryos (n = 180 half-myotomes), dag1 morphants (n = 254 half-myotomes), and dag1 morphants receiving NAD+ (n = 249 half-myotomes) at 6 hpf. NAD+ treatment made no significant difference. (F) MTJ angle quantification. MTJ angles are significantly reduced in dag1 morphants receiving NAD+ (n = 98 MTJs) compared to untreated dag1 morphants (n = 97 MTJs). (G) Bar graph of the percent skeleton length of control embryos per myotome. Note that the average percent skeleton length is more drastically reduced in fkrp morphants (51.4%, n = 153 half-myotomes) than in dag1 morphants (83.9%, n = 260 half-myotomes). NAD+ supplementation increased this percentage in fkrp morphants (61.9%, n = 183 half-myotomes) but has little effect on dag1 morphants (85.0%, n = 252 half-myotomes) Scalebars are 50 μm, error bars in (G) are standard error of the mean. *p < 0.05, **p < 0.01, ***p < 0.001, ns non-significant