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. 2021 Apr 21;9:623394. doi: 10.3389/fcell.2021.623394

FIGURE 2.

FIGURE 2

Characterization of Nsun5 knockout mouse model and OFT septation development delay shown in Nsun5 absent mice. (A) Two sgRNAs designed to target Exon3 of Nsun5 gene. The PAM sequences were highlighted in green and the sgRNAs sequences were highlighted in red. (B) Genotype identification of offspring mice derived from founder mouse with 22-bp deletion. Nsun5 DNA were from tail biopsies of wild-type (+/+), heterozygous (±) and homozygous (–/–). (C) Sanger sequencing verification of offspring mice. (D) Western blot analysis of Nsun5 in mouse heart tissue protein extracted at E14.5. (E) Frequency analysis of OFT development delay in E14.5 embryo hearts. (F) Representative H&E staining of E14.5 hearts. Nsun5+/– and Nsun5– /– mice E14.5 hearts clearly showed OFT aorta misalignment (asterisk) and septation defect (arrow). Scale bar = 100 or 50 μm. Pearson’s Chi-squared test was used for statistical calculation.