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Fig. 4 Phenotype caused by CPA inhibition. (A-C) At 3.5 and 5.3 hpf, the external yolk syncytial layer (e-YSL) could be observed in control embryos. (D-F) Embryos treated with 3′-dA beginning at the one-cell stage to 3.5 hpf still had visible e-YSL at 3.5 hpf, but this structure disappeared by 5.3 hpf, and epiboly did not proceed further (n=5). (G-J) Whole-mount in situ hybridization with the YSL marker camsap2a2 confirmed the absence of e-YSL up to 8 hpf (n=8). Other gastrulation markers, including dorsal shield (K-L; n=8) and mesendodermal cells expressing wnt8 (M,N; n=8), were also absent.
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