DEV02400 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. The Drosophila sog null mutation has a minor effect on the development of ventral structures. (A,B) Expression of sim mRNA in wild-type (A) and sog6 mutant (B) (François et. al., 1994) embryos at stage 10. (C,D) Expression of Pros protein in wild-type (C) and sog6 mutant (D) embryos at stage 11. The monoclonal antibody MR1A was used to detect the Drosophila Pros protein (Spana and Doe, 1995). All embryos are viewed from the ventral side. Anterior is towards the top. Scale bar: 100μm.

• Supplemental Figure 2 -

  Fig. S2. Flat preparation of a twinned embryo of the spider Achaearanea tepidariorum that arose spontaneously. The embryo was stained for DNA. The twins are connected at their caudal regions. This spider species, like other spider species previously examined, can develop a secondary embryonic axis. a, anterior; p, posterior.

• Supplemental Figure 3 -

  Fig. S3. Molecular phylogenetic analyses of At-Sim and At-Omb based on the neighbor-joining method (Saitou and Nei, 1987). (A) A tree constructed using regions spanning the bHLH, PAS-A and PAS-B domains of bHLH-PAS family proteins. This tree reveals that At-Sim is more closely related to Sim subfamily members, including Drosophila and vertebrate Sim, than to Drosophila Trachealess (Dm-trh), which belongs to another subfamily. (B) A tree constructed using the T-domains of Tbx family proteins. At-Omb is more closely related to Drosophila Omb and vertebrate Tbx2 and 3 than to other Tbx subfamily proteins. Numbers indicate bootstrap values. Species: Ag, Anopheles gambiae; Bf, Branchiostoma floridae; Ci, Ciona intestinalis; Dm, Drosophila melanogaster; Dr, Danio rerio; Mm, Mus musculus; Sp, Strongylocentrotus purpuratus. Accession numbers of the proteins used are as follows: Ag-sim, XP_319213; Bf-sim, CAD44626; Ci-sim, ci0100133264; Ci-trh, BAE06738; Dm-sim, AAC64519; Dm-trh, AAA96754; Dr-sim1, AAK27261; Dr-Sim2, AAH65360; Dr-nPAS3, XP_687851; Mm-Sim1, BAA28270; Mm-sim2, NP_035507; Mm-nPAS1, NP_032744; Mm-nPAS3, NP_038808; Sp-sim, XP_782984; Bf-Tbx1/10, AAG34887; Bf-Tbx2/3, AAG34888; Bf-Tbx6/16, AAG34890; Dm-Ombr1, CAA76529; Dm-Omb, AAA28736; Dm-Doc, BAA87864; Dr-Spt, AAC28848; Dr-Tbx2, AAF59835; Dr-Tbx4, AAF59836; Dr-Tbx5, AAF59837; Dr-Tbx6, P79742; Mm-Tbx1, AAG61088; Mm-Tbx2, NP_033350; Mm-Tbx3, AAH96551; Mm-Tbx4, CAI25365; Mm-Tbx5, NP_035667; Mm-Tbx6, P70327.

  Additional references

  Saitou, N. and Nei, M. (1987). The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4, 406-425.

  Spana, E. P. and Doe, C. Q. (1995). The prospero transcription factor is asymmetrically localized to the cell cortex during neuroblast mitosis in Drosophila. Development 121, 3187-3195.

• Movie 1 -

  Movie 1. Time-lapse observation of an At-sog RNAi embryo (left) and a normal embryo (right). The embryos were dechorionated with bleach, attached to the bottom of dish by double-sided sticky tape and overlaid with halocarbon oil 700 (Sigma). A stereo microscope (SZX12, Olympus) equipped with the ORCA-3CCD camera system (Hamamatsu Photonics) was used for observation. Images taken every 15 minutes were edited using Aquacosmos software (Hamamatsu Photonics) and Adobe Premiere to produce this movie. Time (hours) is shown at the left side of the bottom in each panel.