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. 2014 Sep 30;9(9):e107041. doi: 10.1371/journal.pone.0107041

Figure 1. Analysis of DNA, protein, and morphological defects in the l11Jus8 mutant mouse.

Figure 1

(A) Three coding mutations found in the 35 Mb candidate intervals on mouse chromosome 11 are located in the Med31, Nf1 and Erbb2 genes. Only recombinant embryos with the C57BL/6 homozygous DNA fragment inherited from the mutagenised parent which contained the Erbb2 mutation were lethal at weaning, whereas embryos with homozygous Med31 and Nf1 and heterozygous Erbb2 mutations were viable. Straight and dotted lines depict homo- and heterozygous DNA, respectively. (B) Confirmation of the T98,433,986G base change in the Erbb2 gene in l11Jus8 mutants by Sanger sequencing. (C) Phenotype of wild type, homozygous l11Jus8 and Erbb2M802R mutant embryos and hearts. Scale bars: 2 mm for embryos, 500 µm for hearts. (D) Erbb2 protein structure: FLD - Furin-Like Domain, STKD - Serine/Threonine-Tyrosine Kinase Domain. Partial sequence of the STK domain (60 amino acids around the mutation point) is shown for mouse (M), human (H), chick (C) and zebrafish (Z) Erbb2 protein. Asterisks indicate conserved amino acids, colon and period indicate conservation between groups of strongly and weakly similar properties, respectively. Arrowhead demarcates the location of the amino acid change. (E) Predicted changes in the protein structure of the Erbb2M802R conserved kinase catalytic domain based on the reported crystal structure of the Erbb2 protein. (F) Top panel: expression of the Erbb2 mRNA in atria (A) and ventricles (V) of hetero- and homozygous l11Jus8, and embryonic erythrocyte (blood) samples. Prpf8 mRNA expression is used as a positive control for cDNA isolation in blood sample. Bottom panel: Erbb2 expression in the yolk sack (YS) of homozygous l11Jus8 embryo, “-RT” control, genomic DNA (gen) control. A no template negative control (neg) is shown in both panels. (G) Erbb2 immunohistochemistry in heterozygote and l11Jus8 atria (Atr) and ventricles (Ven). Embryonic erythrocytes within the cardiac chambers are indicated with an arrowhead. Scale bar  =  200 µm.