Generation of phd3-null zebrafish and phenotype analysis.A, schematic of the sequence information of phd3-null zebrafish. 8-bp nucleotides (5′-GGAGAGTT-3′) were deleted in exon 1 of phd3-null mutant, resulting in a reading frame shift. B, the predicted protein products in the mutants (70 aa) and their wildtype siblings (243 aa). aa, amino acids. C, verification of CRISPR/Cas9-mediated zebrafish phd3 disruption by heteroduplex mobility assay (HMA). D, qPCR analysis of phd3 mRNA in phd3-null zebrafish larvae and their wildtype siblings (phd3−/− or phd3+/+) (3 dpf). E, photographs of phd3-null zebrafish larvae and their wildtype siblings (phd3−/− or phd3+/+) (3 dpf or 6 dpf). Scale bar = 1 mm. F, measurement of body weight of phd3-null adult zebrafish and their wildtype siblings (phd3−/− or phd3+/+) (4 mpf). G, measurement of body length of phd3-null adult zebrafish and their wildtype siblings (phd3−/− or phd3+/+) (4 mpf). ns, not significant; ∗∗∗∗p < 0.0001, using unpaired Student's t test; data are representative of three independent experiments (mean ± SD of three technical replicates).
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