Fig. 1. Targeted disruption of the Mili gene. (A) Schematic representation of the wild-type allele, the targeting vector and the mutated alleles. The numbered boxes (1-6) denote the 5'-end non-coding exon (exon 1) and the five coding exons (exons 2-5). The targeting vector includes the PGK-neo gene (neo) and the thymidine kinase gene (TK). (B) Southern blot analysis of representative offspring from heterozygous mating. The wild-type allele produces a 10.1 kb EcoRV product, while the disrupted allele gives rise to a 5.5 kb band with the 5'-end hybridization probe. (C) Western blot analysis of the MILI and MIWI proteins from testes using antibody 26F, which recognizes both MILI and MIWI. Lysates (10 µg of protein in each lane) of the testes were loaded on the gel. The +/+, +/- and -/- designations indicate samples from wild-type, heterozygous and homozygous mutant testes, respectively. The 293T cells that were transfected with Mili- or Miwi-expressing plasmids and mock plasmids are shown as the controls.