Morpholino knockdown of atrn leads to epiboly delay. (A) atrn mRNA expression pattern in wild-type embryos was detected by whole-mount in situ hybridization at indicated stages. (B, C) Semi-quantitative RT-PCR detection of zebrafish atrn mRNA level at indicated stages. The relative expression ratio was calculated from the band intensity between atrn and β-actin (internal control). (D) Effectiveness of atrn morpholino. 50 pg atrn-5'UTR-GFP plasmid DNA was injected alone or with 10 ng std-MO or atrn-MO at 1-cell stage. The injected embryos were observed for GFP expression at late gastrulation stages. (E) Morpholino knockdown of atrn results in epiboly delay in a dose dependent manner. Embryos were observed and imaged at indicated stages. Magnified observations of the embryonic marginal cell layers are shown at the last panel. (F, G) The quantitative data of epiboly initiation at 5 hpf and epiboly progression at 8 hpf in (E) are shown. *, p<0.01. Scale bars: 100 μm in (A, E); 500μm in (D).
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