FGF8 rescues rRNA transcription in ncl?/? embryos. (A) qPCR for 5?ETS, ITS1, ITS2 and 18S in untreated and FGF8-treated ncl+/+ and ncl?/? zebrafish (n=10 per sample) indicates rescue of pre-RNA transcription in FGF8-treated ncl?/? zebrafish at 36?hpf. (B) TUNEL staining of untreated and FGF8-treated ncl+/+ and ncl?/? zebrafish at 28?hpf indicates reduced TUNEL+ cells in FGF8-treated ncl?/? embryos compared with untreated ncl?/? embryos (n=15). (C) qPCR for bmp2 in 36?hpf ncl+/+ and ncl?/? embryos (n=10 per sample) that were untreated or treated with 1?µg/µl FGF8 indicates significant downregulation in bmp2 in untreated ncl?/? embryos and significant upregulation in FGF8-treated ncl+/+ embryos. In FGF8-treated ncl?/? embryos, the bmp2 transcript levels were rescued and comparable with untreated ncl+/+ embryos. actb was used as a housekeeping control. (D) Skeletal preparations of 5?dpf ncl+/+ and ncl?/? larvae that were untreated or treated with FGF8, BMH21, FGF8+BMH21 or BMP2. Exogenous FGF8 and BMP2 treatment rescued the cranioskeletal phenotype of ncl?/? larvae (n=45). All experiments were performed three times. Scale bars: 100 µm (B); 70?µm (D). Data are represented as meanħs.d. *P<0.05 (two-tailed, paired Student's t-test).
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