Fig. 1. Identification of zebrafish amotl2 and sequence alignment of Amotl2 proteins. (A) Schematic illustration of procedures for identification of Fgf-responsive genes. One-cell embryos were injected with 10 pg fgf17b mRNA or 100 pg XFD mRNA that encodes a dominant-negative form of Xenopus Fgfr1. The injected embryos and uninjected embryos were collected at the shield stage for total RNA extraction. mRNAs were isolated from the total RNAs and used for making fluorescent probes. The fluorescent probes were hybridized to cDNAs arrayed in slides. A cDNA was regarded as an Fgf-responsive gene if the signal ratio between fgf17b-injected and wild-type embryos or between XFD-injected and wild-type embryos is greater than twofold. (B) Sequence alignment of human, mouse and zebrafish Amotl2 proteins. The number on the right of each line indicates the position of the last residue on the line. The coiled coil domains are underlined with thick lines; the glutamine-rich domain is underlined with a thin line; and the PDZ-binding domain at the C-terminus is boxed. The conserved residues were shadowed. Human AMOTL2 (hAMOTL2) and mouse Amotl2 (mAmotl2) sequences were derived from GenBank with accession numbers NP_057285 and Q8K371, respectively.