Expression of human MUT AP1G1 mRNA resulted in a strong reduction in nrd1-positive motoneurons in 48 hpf embryos. (A) nrd1-dependent EGFP fluorescence in zebrafish embryos after injection of either WT or MUT AP1G1 mRNAs. Representative images (obtained with a fluorescence microscope) of three independent experiments with 25 embryos per condition in each replicate (total number of embryos with phenotypes is shown in red numbers for each experimental condition in panel (B). Lateral views of the head and spinal cord regions (20× magnification) are shown, with further magnification (40×) for the region of the spinal cord in the inset in three distinct experiments and in 10 embryos per treatment (a total of 30 embryos for each experimental condition). (B) Lateral view of a segment of the spinal cord from the WISH experiment performed with the ngn1 probe on zebrafish embryos after injection of either WT or MUT AP1G1 mRNAs. (C) Violin plot distribution of ngn1-positive neurons. The central dashed line marks the median (Q2), and the superior and inferior dotted lines mark the third (Q3) and first (Q1) quartiles. To quantify the number of neurons, statistical significance among and between groups was calculated using one-way ANOVA with post hoc Tukey’s multiple comparison test (**** = p < 0.0001).
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