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p53 expression in the vhl–/–; vll–/– double mutants is downregulated in comparison to that in their siblings. (A and B) Embryos from vhl+/–; vll–/– pair mating were collected and irradiated at 2dpf with γ-ray at a dose of 20 gray. q-PCR was performed 24 hours post γ-ray to quantify the full length of p53 and short form of Δ113p53. This revealed that there was a significant decrease in both forms of p53 expression in the vhl–/–; vll–/– embryos in comparison to their siblings. ****p < 0.0001, two way ANOVA test. The statistical analysis was performed with ddCt values. (C) We also treated embryos from vhl+/–; vll–/– incross with 10 nM CPT and both isoforms of p53 induction was examined in the vhl–/–; vll–/– embryos and in their siblings by Western blot. After treatment, there was a clear reduction in both isoforms of p53 expression in the vhl–/–; vll–/– embryos in comparison to their siblings.
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