The generation of Mtl5 knockout (<i>Mtl5</i>^<i>-/-</i>) mice.

(A) Diagram (top) and nucleotide sequence (bottom) of mouse Mtl5 locus taken from the Ensemble database (ENSMUST00000025840.15) with exons (green boxes) and TGA (red) premature stop codon of the modified transcript. CF and CR represent the genotyping primers of Mtl5 knockout mice. PAM, protospacer adjacent motif. sgRNA, single-guide RNA. A mouse strain with a 19 bp deletion starting from position 498 to 516 in exon 2 was confirmed by PCR (B) and Sanger sequencing (C). (D) Western blot analysis of MTL5 expression in testes of 10-week-old mice with corresponding genotypes. (TIF)