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Fig. 1
Generation of the cyp4v7 and cyp4v8 double knockout zebrafish. (A) Phylogenetic relationships and gene structures of CYP4 family genes in human and zebrafish. (B) In situ hybridization of cyp4v7 and cyp4v8 in retinas of two-month-old WT zebrafish. IS+OS, inner segment and outer segment; ONL, outer nuclear layer; INL, inner nuclear layer; GCL, ganglion cell layer. Scale bars: 20 µm. (C) Semiquantitative RT-PCR analysis of cyp4v7 and cyp4v8 expression in two-month-old zebrafish eyes. (D) CRISPR/Cas9 target sites. (E) DNA sequencing of the corresponding genomic region for WT and mutant zebrafish. The sequencing traces revealed a 2 bp deletion/6 bp insertion mutation in cyp4v7 and a 2 bp deletion mutation in cyp4v8. (F) Relative mRNA levels of cyp4v7 and cyp4v8 were detected by qPCR in WT and cyp4v7 and cyp4v8 double knockout zebrafish at 1 mpf. The results are shown as mean ± SD. ***P < 0.001.
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Full text @ Invest. Ophthalmol. Vis. Sci.