Transcriptional consequences of dhps morpholino (MO) and eif5a1/2 MO in zebrafish. RNA sequencing was performed on RNA isolated from zebrafish embryos. At 24 h post fertilization, control MO was injected at 8 ng/embryo, dhps MO was injected at 4 ng/embryo alongside 4 ng/embryo of control MO, and eif5a1 and eif5a2 MOs (eif5a1/2 MO) were coinjected at 4 ng/embryo each. N = 3 independent biological samples of 25 embryos each, and RNA was harvested at 3 days postfertilization. A, principal component analysis of altered gene expression with dhps or eif5a MO in zebrafish embryos. Black dashed circles represent control zebrafish, red dashed circles represent dhps MO zebrafish. B, volcano plot of altered genes in dhps MO embryos compared with control MO embryos; red circles indicate genes that satisfy the cutoff threshold of p-adj <0.05 and fold change> 2. C, volcano plot of altered genes in eif5a MO embryos compared with control MO embryos; blue circles indicate genes that satisfy the cutoff threshold of p-adj <0.05 and fold change >2. D, relative expression levels of dhps, eif5a1, and eif5a2 from the RNA sequencing data from the indicated MO datasets. E, Venn diagram of the overlap between genes altered by dhps MO versus control MO and genes altered by eif5a MO versus control MO. F, heat map showing the fold change of the 20 genes altered in both dhps MO versus control MO and eif5a MO versus control MO. G, Gene Ontology (GO) pathways altered by dhps MO versus control MO. H, Reactome pathways altered by dhps MO versus control MO. Data are presented as mean ± SEM, and statistical significance was determined by p-adj <0.05. eif5a, eukaryotic translation initiation factor 5A.
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