DEV02776 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Rho specificity is conserved in zebrafish Arhgef11. Comparison of the indicated amino acids within the DH domains of human and zebrafish Arhgef11 with the human residue numbers indicated above the alignment. Highlighted are the identical (yellow) and similar (green) amino acids required for Rho interaction and specificity.

• Supplemental Figure 2 -

  Fig. S2. Endoderm and dorsal forerunners cells, which ultimately form Kupffer’s vesicle, appear normal in morphants. In situ hybridization using sox17 probe on 8.5 hpf uninjected WT (A,A′) and MO^AUG-injected (B,B′) embryos demonstrates formation of endoderm and dorsal forerunner cells (arrows) is not disrupted in morphants. Embryo length (marked by arrowheads) is also unaffected. Probe used was as described in Alexander and Stainier (Alexander and Stainier, 1999).

• Supplemental Figure 3 -

  Fig. S3. Midline structures and patterning are not impaired by Arhgef11 loss-of-function. In situ hybridization using olig2 probe on cryosections of WT control (A) and MO^AUG-injected (B) embryos indicates normal patterning of the neural tube (outlined by white dotted lines). Notochord also appears normal (outlined by black dotted lines). In situ hybridization using probe for the Hedgehog target gene, patched1, on WT (C,C′) and morphant embryos (D,D′) also reveals no abnormalities. Probes for olig2 and patched1 were as described by Park et al. (Park et al., 2002) and Concordet et al. (Concordet et al., 1996), respectively.

• Movie 1 -

  Movie 1. Cilia in the pronephric duct lumen of WT control embryos at 2.5 dpf acquired at 250 frames per second. The portion shown here is 1 second slowed to 15 frames per second. The cilia beat an average of 25-26 times in one second, equal to a frequency of 25-26 Hz.

• Movie 2 -

  Movie 2. Cilia in a cystic region of the pronephric ducts in MO^AUG-injected embryos at 2.5 dpf. Images were acquired and processed as in Movie 1. The cilia beat an average of 35-36 times per second, equal to a frequency of 35-36 Hz.

  References

  Alexander, J. and Stainier, D. Y. (1999). A molecular pathway leading to endoderm formation in zebrafish. Curr. Biol. 9, 1147-1157.

  Concordet, J. P., Lewis, K. E., Moore, J. W., Goodrich, L. V., Johnson, R. L., Scott, M. P. and Ingham, P. W. (1996). Spatial regulation of a zebrafish patched homologue reflects the roles of sonic hedgehog and protein kinase A in neural tube and somite patterning. Development 122, 2835-2846.

  Park, H. C., Mehta, A., Richardson, J. S. and Appel, B. (2002). olig2 is required for zebrafish primary motor neuron and oligodendrocyte development. Dev. Biol. 248, 356-368.