Targeted mutation of Adam11. (A) Top; Domain structure of ADAM11 protein. PRO, proprotein; MP, metalloprotease-like; DIS, disintegrin-like; CR/EGF, Cysteine-rich and EGF-like repeat; TM, transmembrane; Cyto, cytoplasmic domain. Adam11 gene was disrupted by the insertion of a termination codon and a PGKneo cassette into exons 5 – 7. An MC1/TK cassette at the end of the targeting vector allowed for negative selection. The BE2K-probe used for Southern blot analysis, and the expected BamHI "B" fragments of the targeted allele (TG) and wild-type allele (WT) are indicated by arrows. The primers, AGN1 and 033 were used for the selection of the recombinant ES clones. (B) Southern blot analysis of mouse genomic DNA. The expected DNA fragments for the wild-type allele (WT) and targeted allele (TG) are 13.7-kb and 10.3-kb, respectively. +/+, wild-type; +/-, heterozygote; -/-, homozygote. (C) Western blot analysis of ADAM11 expression in the mouse cerebellum. Absence of ADAM11 protein in the (-/-) mutant cerebellum was shown using anti-ADAM11 monoclonal antibody. "P" indicates the positive control, HA-tagged mouse ADAM11 protein expressed in HeLa cells. White circle indicates mature-form of ADAM11 protein and black circle shows immature-form, which was not processed. (D) Hippocampus sections with HE stain. Scar = 30 μm. (E) Cerebellum sections with HE stain. Scar = 200 μm.