Fig. 2
From: Hepatic lipid metabolism is altered in Ubiad1+/− mice of both sexes
Generation of Ubiad1+/- mice by the CRISPR Cas9 system. (A) A schematic drawing of the Ubiad1 locus, binding sites for the two sgRNAs, and the generated disrupted Ubiad1 allele. The 29 base pair deletion generating a frameshift mutation in the Ubiad1 gene is marked on the WT allele. Amino acids marked with red color show the expected codon alterations generated due to the introduced frameshift mutation. (B) Migration of DNA fragments obtained with PCR amplification of a normal and mutated Ubiad1 alleles. The predicted sizes of the DNA fragments are 265 bp for the Ubiad1+/+ allele and 236 bp for the Ubiad1+/- allele. The gel image was cropped and original gels are presented in Supplementary Figure 5A. (C, D) Quantification of wild type and mutated Ubiad1 mRNA transcripts, versus only wild type mRNA in (C) subcutaneous WAT and (D) liver in 20 weeks old mice fed HFD for 10 weeks. (n = 11−13 per group in males, n = 9 per group in females). Data were normalized against the expression of Tbp and Rplp0, respectively. (E) Western blot of Ubiad1 and beta-actin on whole liver proteins from Ubiad1+/+ and Ubiad1+/- mice (n=6 per group). One sample from Ubiad1+/+ male was excluded from the quantification analysis due to saturated band. The blots were cropped and original blots are presented in Supplementary Figure 5B. Data represent means ± SD. P-values were calculated using unpaired t-test. *p < 0.05, **p < 0.01, ***p< 0.001.
