DEV01257 Supplemental Figures

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Schematic of the wild-type Disp1 allele (Disp1^wt), the conditional allele of exon 2 (Disp1^D2c) and the allele with exon 2 deletion (Disp^D2). The locations of the fragments used as Southern hybridization probes are shown. 5¢ probe detects an 11 kb SacI fragment from the wild-type allele and a 7 kb fragment from mutated allele. 3¢ probe detects a 7 kb KpnI fragment from wild type and a 9 kb fragment from mutated allele. After ubiquitously expressed Cre-mediated recombination (by crossing with b-actin-Cre transgenic line), Disp^D2 allele was generated and confirmed with Southern hybridization in which exon 2 is absent from Disp^D2/D2 animals.

• Supplemental Figure 2A
• Supplemental Figure 2B -

  Fig. S2. (A) Protein sequence alignment of Disp1, Disp2 and Drosophila Disp. The conserved regions are marked by a green background. The identical amino acids in all three sequences are highlighted in blue. Residues with greater than 50% similarity are black on a green background. Identical residues in only two of the sequences are highlighted in red. Weakly similar residues with 20% similarity are in purple. Predicted transmembrane domains are overlined. A blue line indicates the intron/exon boundary. (B) Schematic of alternative splicing in Disp1^D2/D2 mutant. In wild type, exon 1B is joined by exon 2 (with starting ATG), followed by exon 3. The predicted translational product is 1521 aa. In Disp1^D2/D2 mutant, exon 1A is joined by exon 3, followed by exon 4 (containing alternative starting ATG). The predicted N-terminal truncated Disp1^D2 is 1328 aa.

• Supplemental Figure 3 -

  Fig. S3. Midline patterning defects in Disp1^D2/D2 and Disp1^D2/C829F mutants. (A,D,G) wild-type animals; (B,E,H) Disp1^D2/D2 mutants; (C,F,I) Disp1^D2/C829F mutants. (A-C) Histological sections through upper jaw reveal the loss of primary palate and upper incisors in Disp1^D2/D2 and Disp1^D2/C829F mutants. (D-F) Midline structure defects in nasal septum. Note the midline localization of vomeronasal organ in Disp1^D2/D2 mutant and the absence of this structure in Disp1^D2/C829F. (G-I) Pituitary hypoplasia in Disp1^D2/D2 mutant. Note that no pituitary develops in Disp1^D2/C829F mutant. UI, upper incisor; PP, primary palate; VN, vomeronasal organ.