Figure 1

Alerted gene expression in gankyrin-injected zebrafish embryos. (A) Genes involving zf p53, zf p21 and Mdm2 were analyzed using single-embryo RT-PCR in embryos injected with pCS2+EGFP hgankyrin and buffer. Zf p53 and zf p21 expression was lower in embryos injected with hgankyrin compared to those injected with buffer as negative control. There was no difference in zf Mdm2 expression between hgankyrin injection and buffer, suggesting that hgankyrin regulates the expression of p53 but not Mdm2. (B) Zf p53 mRNA expression between injected with pCS2+EGFP hgankyrin and buffer was analyzed by RT-PCR. The results are determined as average ratios of the OD of zf p53 and to the OD of β-actin (average and standard errors of the three experiments, P value < 0.05). Above the results (Figure 1A), level of zf p53 only decreased in hgankyrin injection embryos. (C) Northern blot analysis of p53 and Mdm2 expression following hgankyrin overexpression in zebrafish. Total RNA were prepared at 24 hpf of each lane and distributed by formaldehyde-agarose gel electrophoresis. 15 µg of RNA were loaded in each lane and hybridized using p53 and Mdm2 probes. The p53 levels are down regulated in hgankyrin injected embryo at 48 hpf. The loading control for lanes serves 18S rRNA band photographed from the etiethidium bromide stained gel.