Fig 8

A. Illustration showing foxo3b gene structure in indicated vertebrates. The distance between the stop codon and the proximal 3′UTR intron is on the right. Open rectangles: UTRs, filled rectangles: coding region, gray lines: introns (hash marks denote shortened intron sequences). B. Top: Western blot showing protein levels in wild-type sibling (lane 1) and magoh mutant (lane 2) embryos at 21 hpf. Bottom: a dot plot showing Foxo3b levels normalized to tubulin levels in magoh mutant embryos and WT siblings at 21 hpf in three biological replicates. (N = 5 embryos per genotype per replicate). Error bars: standard error of means. C-F. Confocal images showing Myh1 immunofluorescence using anti-A4.1025 in somites 12–16 of WT sibling (C), magoh^-/- mutant (D), magoh^-/-; foxo3b^+/- mutant (E), and magoh^-/-; foxo3b^-/- mutant (F) embryos at 26 hpf (N = 13 embryos/genotype). G-J. Merged confocal images showing motor neurons (red; detected by anti-SV2 staining) and acetylcholine receptors (green; detected by alpha-bungarotoxin staining) in somites 12–16 of WT sibling (G), magoh^-/- mutant (H), magoh^-/-; foxo3b^+/- mutant (I), and magoh^-/-; foxo3b^-/- mutant (J) embryos. Neuromuscular junctions in the merged images are yellow. White arrowheads point to the distal end of the motor neuron. (N = 13 embryos per genotype). Scalebar in J (for panels C-J) is 100 nm. K. Boxplots showing quantification of motor axon length in embryos of genotypes indicated along the x-axis (4 motor neurons/embryo and 13 embryos/genotype). Welch’s t-test p-values for comparison between magoh^-/- mutant and magoh^-/-; foxo3b^-/- mutant embryos are at the top.