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Fig. 2 Reporter mRNA with a shortened pou5f3 3′UTR is effectively translated in the early development of zebrafish. (A) Schematic diagrams of reporter genes with the SV40 3′UTR (SV40), full-length pou5f3 3′UTR (Long), and processed pou5f3 3′UTR (Short). (B) Left: Whole-mount in situ hybridization of reporter mRNAs in transgenic zebrafish embryos at 2 hpf with the GFP antisense probe. Insets are embryos hybridized with the GFP sense probe. bd, blastodisc. Similar results were obtained from two lines of transgenic zebrafish. Right: Reporter mRNA levels were determined by quantitative RT-PCR (means ± SD; n = 3). (C) Detection of newly synthesized GFP using Puro-PLA in embryos at 2 hpf. DNA is indicated in blue. Scale bars, 100 μm (B) and 10 μm (C). (D) Number of Puro-GFP PLA sites per 40,000 μm2 was counted (means ± SD; n ≥ 18). Similar results were obtained from three independent experiments. **P < 0.01 and *P < 0.05, Mann-Whitney’s U test.