Figure 3. Mutation of nucleoredoxin in l11Jus13 (Nxn^J13/J13).

(A) Haplotype map of 239 unaffected and 83 affected mice used for meiotic mapping. The location of each marker and Nxn is displayed (Ensembl v52). The mutation lies in a 6Mb region located between rs3702197 and rs13481117. (B) The Nxn locus is depicted with introns 6 and 7 boxed. (C) The boxed region is expanded to illustrate the consequence of splicing in the wild type and mutant. A transversion (T to A) abolishes a consensus splice donor site leading to aberrant RNA splicing. The six base pair cryptic splice site used in the mutant is underlined. (D) Aberrant splicing in Nxn^J13/J13 predicts an in frame insertion of 10 amino acids, GMELEGKWKA, (white), which occurs within the acidic region (black) of Nxn. (E) RT–PCR using primers flanking exon 6–7 from pools of three E14.5 heads of wild-type, heterozygous, and homozygous mutants demonstrates aberrant splicing in the mutant allele. (F) Western blots of Nxn and Actin from wild-type, heterozygous and homozygous mutants at E12.5 show reduced protein in the homozygous mutant. A reduction in protein was also observed at E15.5 and E18.5, and a polyclonal antibody against the N-terminus gave similar results (data not shown).