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Fig. 1 psmb6 mRNA expression pattern and generation of psmb6 mutants. (A) ISH for psmb6 at 24 hpf, 36 hpf, 48 hpf, and 3 dpf. (B) Knockout of psmb6 by CRISPR-Cas9 resulted in two different types of psmb6 mutants, with deletions of 4 bp and 42 bp (five bases are located in exon 3, and thirty-seven bases were located in intron 3–4). Sanger sequencing confirmed the 4-nt and 42-nt deletion mutations. Detection of mutant knockout efficiency was performed by Western blotting. (C) psmb6 mutants displaying cerebral hemorrhage (red arrow) and cerebral necrosis (blue arrow). Scale bars: 100 μm. (D) Phenotypic rates of two types of psmb6 mutants at 4.5 dpf. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)