Fig 2. Generation of A/J-12^SM Iah1 knockout (KO_A12) mice.

(A) The sequence of crRNA (boxed) which targets the Cas9 nuclease to a region of exon 1 in the Iah1 mouse gene. The mouse with a 53bp deletion and a frameshift mutation was identified. (B) Real-time qPCR analysis of Iah1 mRNA levels in WT_A12 and KO_A12 mice. Data were expressed as mean ± SEM. WT_A12 (n = 4), KO_A12 (n = 4). **, Significantly different (p<0.01) when compared with WT_A12 mice by student’s t-test. (C) Western blot analysis of the IAH1 protein (10 μg each, 28 kDa) with α-Tubulin as a loading control in WT_A12 and KO_A12 mice. The size marker (Precision Plus Protein Standards, Bio-Rad) was loaded in lane M. Tissues were collected from WT_A12 and KO_A12 mice fed with a high-fat diet for 12 weeks. BAT, brown adipose tissue; eWAT, epididymal white adipose tissue.