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Files in this Data Supplement:
Fig. S1. Quantification of the number of rosettes in fgf3;fgf10 double mutant primordia at different time-points during lateral line development. At 28 hpf, all double mutant embryos show a complete absence of rosettes (blue bar, n=12), rather than the normal set of 3-4 that can be observed in wild type. However, the number of rosettes gradually increases, such that by 42 hpf almost half of the embryos display the normal number (green).
Fig. S2. Expression of fgf ligands in fgf3;fgf10 double mutants. (A,B) The lower panels are transmission pictures, upper panels are merge of the GFP channel and the transmission channel after colour inversion (see Material and methods). fgf3 (A) and fgf10 (B) are expressed strongly and uniformly throughout the primordia.
Fig. S3. Time-lapse analysis showing epithelialized cells snapping together to form rosettes in the second phase of washout. The embryo was pre-treated with SU5402, washed for 5 hours and mounted side-on. Asterisks indicate the formed rosettes (see also Movie 7).
Movie 1. Time-lapse movie of a wild-type primordium from 30 hpf to 36 hpf. The highly organized and smooth migration of the tissue and the deposition of one neuromast from the trailing edge can be seen. Images captured every 3 minutes using a Perkin Elmer Ultraview LCI spinning disk, 20×/NA0.75 objective. Movie length: 382 minutes.
Movie 2. Time-lapse movie of an fgf3;fgf10 double mutant primordium from 30 hpf to 42 hpf showing repetitive cycles of tissue extension. Images captured every 3 minutes using a Perkin Elmer Ultraview LCI spinning disk, 20×/NA0.75 objective. Movie length: 750 minutes.
Movie 3. Time-lapse movie of a cldnb:gfp embryo in SU5402. The embryo was pre-treated with SU5402 and shows the uncoordinated movement of the tissue, and the back and forth movement of the cells within. Images captured every 3 minutes using a Perkin Elmer Ultraview ERS spinning disk 20×/NA0.75 objective. Movie length: 441 minutes.
Movie 4. Time-lapse movie of the primordium of a cldnb:gfp embryo mounted in 80 µM SU5402 and imaged directly to show the progressive melting of rosettes. The movie begins 30 minutes after the start of exposure to SU5402. Images captured every 5 minutes using a Perkin Elmer Ultraview ERS spinning disk 20×/NA0.75 objective. Movie length: 380 minutes.
Movie 5. Time-lapse movie of an SU5402 washout experiment showing the reassembly of rosettes followed by the resumption of migration. Movie starts after 2 hours of washes. Images captured every 3 minutes using a Perkin Elmer Ultraview LCI spinning disk 20×/NA0.75 objective. Movie length: 843 minutes.
Movie 6. Time-lapse movie of the primordium of a 22-hpf cldnb:gfp embryo showing the concerted apical constriction of several cells at the back of the leading region to form a new rosette. Migration has not yet started at that stage. Images taken every 3 minutes with a 40× water objective. Movie length: 216 minutes.
Movie 7. Time-lapse movie from a side view of the primordium during a washout experiment showing the reformation of rosettes. The movie was taken during the second phase of washout (see Fig. 5F) and starts after 5 hours of washes. Images were captured every 3 minutes with an Olympus Fluoview 1000 40×W/NA1.2 objective. Movie length: 594 minutes.
Movie 8. Time-lapse movie of a wild-type mosaic primordium exposed to DMSO showing lynTdtomato-expressing cells transplanted into a cldnb:gfp primordium all projecting in the direction of migration. Images captured every 90 seconds using a Perkin Elmer Ultraview ERS spinning disk 40× objective. Movie length: 69 minutes.
Movie 9. Time-lapse movie of a wild-type mosaic primordium exposed to SU5402 showing lynTdtomato-expressing cells transplanted into a cldnb:gfp primordium. In the absence of FGF activity, the cells form many more filopodia than do controls, which protrude in all directions, attesting a mesenchymal-like phenotype. Images captured every 90 seconds using a Perkin Elmer Ultraview ERS spinning disk 40× objective. Movie length: 180 minutes.
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