DEV01374 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. A single full-length zebrafish met open reading frame was identified by assembling a predicted gene structure from the zebrafish genome, and confirmation that this predicted sequence was expressed was obtained from a combination of cDNA library screening and RT-PCR. Two predicted hgf genes could be identified by mining of the zebrafish genome sequence in a similar fashion. Owing to both incomplete sequence and mis-assembly of the genomic scaffold from which the two different sequences are derived, only a partial sequence could be predicted for both genes. cDNA sequences were isolated by RT-PCR encompassing the genomic fragments and used for further analyses. (A) Alignment of zebrafish met with identified mammalian sequences. Alignment shown is of the region encoding the tyrosine kinase domain of the different Met proteins. (B) Partial alignment of the two predicted HGF proteins identified by mining of the zebrafish genome sequence with known HGF proteins. Owing to both incomplete sequence and mis-assembly of the genomic scaffold from which the two different sequences are derived, only a partial sequence could be predicted for both genes, and only the region of overlap between the encode proteins is aligned. In A and C, identities are shaded (red, full; blue, partial) and alignment gaps are indicated by a broken line. Amino acid alignments were examined and formatted using GeneDoc (version 2.6.02; http://www.psc.edu/biomed/genedoc/).

• Movie 1 -

  Movie 1. Time-lapse of the migration of the PHM primordia, initiating at 48 hpf for 200 minutes. Frames are taken 5 minutes apart using near simultaneous DIC bright field and fluorescent image capture on an a actin GFP transgenic embryo. Frame rate is accelerated to 1200X real time.

• Movie 2 -

  Movie 2. Time-lapse of the tip of the migrating PHM primordia initiating at 60 hpf for 30 minutes. An oblique ventral view of an a actin GFP embryo at 60 hpf, with the distal tip of the PHM in focus centre left and its attachment point at the cleithrum bottom left and the sternohyal muscle out of focus also at the bottom left. Frames are taken 30 seconds apart using near-simultaneous DIC bright-field and fluorescent image capture. Frame rate is accelerated to 150X real time.