Figure 3.

Targeted disruption of the mDido locus. (A) Scheme showing exon-intron structure of the Dido locus. Colors indicate common and specific nucleotide sequences. Noncanonical splicing events are indicated with stars. To target the murine Dido locus, a region containing exon IV and most of exon III was deleted and replaced by a neoresistance cassette. A restriction map is shown of WT mouse Dido. Arrow indicates the position of M423. ERV, EcoRV; ERI, EcoRI. (B) Southern blot hybridization. Mouse Dido disruption by homologous recombination resulted in a 5.5-kb band using EcoRV-digested genomic DNA. (C) mDido transcript expression was analyzed in WT (+/+), Dido+/neo (+/neo), and Didoneo/neo (neo/neo) mouse tissues by hybridization with a probe common to the 3 mDido transcripts. The blot was stained with methylene blue to control RNA loading. (D) mDido3 transcript expression was analyzed on the same blot with an mDido3-specific probe.