No difference in the p53 response was detected in MZ-fam83fa−/− embryos compared with WT. (a) Representative western blot for p53 and β-actin (input) in protein extracts from zebrafish embryos treated with IR at 24 hpf and harvested for protein extraction 2 h (t1) and 10 h (t2) after treatment. (b) Quantification of p53 band density for three independent experiments as represented in (a), normalized to β-actin and expressed as arbitrary units. *p < 0.05, two-way ANOVA with Šídák’s multiple comparison test. n.s. = not significant. Error bars = s.d. (c) qRT-PCR of mRNA extracted from WT or MZ-fam83fa−/− KO embryos as labelled, 2 h following treatment with IR at 24 hpf. qRT-PCR was performed for p53 target genes, including p53 itself, and nrz, the zebrafish bcl−2 homologue. Data represent mean of four biological replicates (data points), each consisting of 25 embryos per line, normalized to 18S and expressed as fold induction relative to untreated WT. (d) Same as (c) except at 10 h following IR treatment. Two-way ANOVA with Tukey’s post-hoc test showed no significant differences between IR-treated WT and MZ-fam83fa−/− KO for any gene tested. Error bars = s.d.
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