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Fig. 2
Zebrafish embryos accumulate Pou5f3 protein during the early stages of development through maintaining the granular structures of pou5f3 mRNA
(A) Time courses of Pou5f3 accumulation in immature (Im) and mature (M) oocytes and in embryos at hours post fertilization (hpf). Ribosomal protein l11 (Rpl11) is a loading control.
(B) The intensities of both Pou5f3 bands were quantified (means ± standard deviations; n = 5). NS, not significant; ∗p < 0.05; ∗∗p < 0.01 (Tukey-Kramer test).
(C and D) Whole mount FISH analyses of pou5f3 mRNA (green) in embryos at 0, 1.5, 3, and 6 h post fertilization.
(C) Upper, lateral views of embryos by fluorescent dissection microscopy. Lower, animal pole views of embryos by confocal microscopy. Insets show bright-field views of live embryos. bd, blastodisc.
(D) Upper, high-resolution confocal images. Lower, enlarged views of the boxed regions in the upper images.
(E) Numbers of granules per 100 μm2 in embryos at 0, 1.5, 3, and 6 hpf were counted (means ± standard deviations). The numbers in parentheses indicate the total numbers of oocytes analyzed. Similar results were obtained from two independent experiments.
(F) Distribution of pou5f3 RNA granules in the cell cytoplasm. DNA is shown in blue. n, nucleus. Bars, (C) 100 μm; (D) upper, 20 μm; lower, 5 μm; (F) 10 μm.
(G) PAT assay for pou5f3 mRNA in embryos at 0, 1.5, and 3 hpf. Similar results were obtained from three independent experiments.