Expression of MUT AP1G1 mRNA affects midbrain–hindbrain boundary formation. Expression of zebrafish neurogenin 1 (ngn1) during early development in the midbrain–hindbrain boundary (mhb) of embryos injected with either human WT or MUT AP1G1 mRNA. All treatments, including the controls (non-injected embryos), were performed in triplicate, with 30 embryos per condition (90 embryos per experimental group). The embryos shown in the figure are representative of all experiments. (A) Dorsal views (anterior to the left) of 16 hpf embryos at 25× magnification. (B) Lateral views (dorsal to the top and anterior to the left, 25× magnification) of 48 hpf embryos. (C) Violin plot distribution of midbrain–hindbrain boundary (mhb) measurements at 48 hpf for non-injected and injected embryos Measurements were performed on digital images using the ImageJ software (Version 1.8.0). The Y-axis shows the mhb length (μM). Measurements were performed on 10 embryos for each of the three independent experiments and for each treatment (a total of 30 embryos for each experimental condition). Inside each violin, the central dashed line marks the median (Q2), and the superior and inferior dotted lines mark the third (Q3) and first (Q1) quartiles, respectively. Measurements were analyzed among and between groups using one-way ANOVA with post hoc Tukey’s multiple comparison test (* = p < 0.05; **** = p < 0.0001). (h, hindbrain; m, midbrain; mhb, midbrain–hindbrain boundary).
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