The sart3 mutation triggers alternative splicing of mdm4 as well as the activation of the p53 pathway.A Overlap of gene expression change (padj < 0.05) with alternative splicing (padj < 0.05) in sart3smu471 mutants from RNA-seq data. The overlapping number is 607. B p53 signaling pathway enrichment by GO term enrichment and KEGG pathway analyses from the 607 overlapped genes. The horizontal axis represents the significance, −log10(p). C Heatmap of p53 signaling pathway-related gene changes exhibited in (B) (data was normalized by Z-score, Z-score was shown on the right). The p53 negative regulator, mdm4, is boxed. Dmdm4 alternative splicing and primers designed for PCR detection of exon 6 skipping. The mdm4 full-length transcript (mdm4-FL) decreases and truncated mdm4 transcript (mdm4-S) increase in sart3smu471 mutants. E Decreased expression of mdm4-FL by RT-qPCR at 2.5–4 dpf sart3smu471 mutants (Student’s t-test, means ± SD, n ≥ 15 per group, ****P < 0.0001). F The procedure for the mdm4 overexpression rescue of reduced HSPCs in sart3smu471 mutants. hs: heat shock. G Partial restoration of cmyb signals in sart3smu471 mutants by mdm4 overexpression (OE). Red arrowheads indicate cmyb signals by WISH in the CHT region of 4 dpf larvae. Scale bars: 200 µm.
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