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Fig. 1
Generation of whsc1 mutant zebrafish with Crispr/Cas9 technology. (A) A diagram showing the structure of WHSC1 protein. (B) design of sgRNA against whsc1 gene. This sgRNA site targets the 15th exon. Target sequence was shown with blue letters, and PAM region was shown in red. Green box marked the reorganization site of BSTX1 enzyme that lies within sgRNA target site, loss of which indicated the presence of indels. (C) Restriction enzyme analysis of the presence of indel mutations in two independent mutant lines. (D) Sanger sequencing results of the whsc1 mutant lines. Arrows indicated the locations of indels. (E) WB analysis of the levels of H3K36Me2 and H3K36Me3 in whsc1 wild-type and mutant embryos. (F) Immunofluorescence analysis of H3K36Me2 level in whsc1 wild-type and mutant embryos. Photos of wildytpe and whsc1 mutant embryos were taken under same exposure time.