Placental distribution of ezrin and generation of <i>Ez</i>^−/− mice.

<p>(A) Schematic representation of the wild-type allele, targeting vector, and targeted allele of the mouse ezrin gene (<i>Vil2</i>). The first ATG codon is located in exon 2. The targeting vector contained the neomycin-resistance cassette (neo) combined with engrailed-2 (En-2)/splicing acceptor (SA)/internal ribosome entry site (IRES)/β-galactosidase (lacZ) on its 5′ side and with a polyadenylation site (PA) on its 3′ side, which was designed to be inserted between exons 2 and 5 in the targeted allele. Southern blot analysis of genomic DNA from <i>Ez</i>^+/+, <i>Ez</i>^+/−, and <i>Ez</i>^−/− mice. The NdeI fragments were detected by the 5′ probe from the wild-type (28 kb) and targeted alleles (9 kb), and the BglII fragment was detected by the neo probe from the targeted allele (8 kb). The XbaI fragments were detected by the 3′ probe from the wild-type (6 kb) and targeted alleles (5 kb). Immunofluorescence micrographs of frozen sections of <i>Ez</i>^+/+ (B) and <i>Ez</i>^−/− (C) placenta at gestational day 18.5 obtained with rabbit polyclonal antibody against ezrin (red), showing complete loss of ezrin in <i>Ez</i>^−/− placenta. Nuclei were stained with DAPI (blue). De, decidua; Jz, junctional zone; Lz, labyrinth zone. Arrowheads indicate trophoblast giant cells. Confocal microscopic images of ezrin in labyrinth area were also shown in right panels (B, C). Scale bars represent 100 µm (left, center) and 20 µm (right).