Figure 11.
Under hypoxia, the expression of zebrafish nip3a in wild-type embryos was inhibited by p53 overexpression and up-regulated by p53 knockdown. (A) Under hypoxia (2% O2, 8 h), the expression of nip3a in wild-type embryos injected with synthesized zebrafish p53 mRNA (200–500 pg/per embryo) was reduced (A3, A4; n=20) compared with the embryos without injection (A1, A2; n=20); the expression of nip3a in wild-type embryos injected p53 morpholino (8 ng/per embryo) was increased (A5, A6; n=20) compared with the embryos without injection (A1, A2); the expression of nip3a in p53 mutant embryos injected with synthesized p53 mRNA was decreased (A9, A10; n=20) compared with embryos without injection (A7, A8; n=20). (B) Semi-quantitative RT–PCR analysis further confirmed the results shown in (A). Data of RT–PCR analysis are reported as mean±s.d. of three independent experiments performed in triplicate; the statistical analysis was performed using GraphPad Prism 5.0 (t-tests). (C) The expression of injected p53 mRNA was confirmed by western blot. (D) The specificity and efficiency of p53-morpholono (p53-MO)-mediated p53 knockdown was confirmed by western blot.