(A) WT zebrafish embryo, (B) vehicle control injected with the same volume of phenol red dye in distilled water as the volume of the mRNAs injected, and (C) rnf152 mRNA injected embryo. Microinjection of rnf152 mRNA (200 pg) into embryos at 1- or 2 cell stages for overexpression of rnf152. All images are in lateral view (n = 3). (D) WT, (E) 5′ mismatch MO, and (F) rnf152 MO that 5 ng of rnf152 morpholino was injected into embryos at 1-cell stage for knockdown of rnf152. All images (D–F) are in lateral view of embryos at 24 hpf. (G–J) Graphs showing diameter of the eyes and width of the neural tubes. (G) Diameter of the eyes, (H) width of the neural tube of WT, vehicle control, and rnf152 overexpressed embryos. (I) Diameter of the eyes, (J) width of the neural tube of WT, 5′ mismatch control, and rnf152 MO. Red arrowheads indicate the rhombomeres (r1–7) while dotted red lines indicate the width of the neural tube. Area of the eyes was marked with red dotted lines. The graphs were obtained using ImageJ, error bars depicting standard errors. Unpaired t-test was executed to obtained p-values, which is p<0.0001 (n=3). Abbreviations: fb- forebrain, hb- hindbrain, mb- midbrain, NT- neural tube, ret- eye, SC- spinal cord, and TB- tail bud. Scale bars A–F: 50 μm.
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