Activation of p53-dependent apoptotic pathway contributed to the defective formation of PA in nup62l-mutants. (A) Partial rescue of morphological defects in PA of nup62l-mutants injected with 5 ng p53-MO as shown by the lengths of white lines. (B) TUNEL assays were performed to assess the apoptosis in pharyngeal region of nup62l- and p53- double inactivated mutants compared to WT and nup62l-mutants at 72 hpf. Embryos were imaged by confocal microscopy. PA region, eyes and optic tectum were labeled. (C) PA sagittal sections at 96-hpf of WT, nup62l-mutants or nup62l-mutants injected with 5 ng p53-MO were analyzed with hematoxylin and eosin staining. The 2nd-4th ceratobranchials (cb) were labeled. (D) Expression levels of genes (tp53bp1, tp53i11a and tp53i11b) implicated in p53-dependent apoptotic pathway were analyzed by qRT–PCR in WT, nup62l-mutants or nup62l-mutants injected with 5 ng p53-MO at 72 hpf. Expression levels were normalized to WT embryos. Data expressed as Mean ± SD were representatives of three independent experiments. ** p < 0.01.
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