Supplemental Figure 1 and Supplemental Movies 1–3.

Files in this Data Supplement:

• Supplemental Figure 1. - Supplemental Fig. 1. Developmental expression of zMPO:GFP. Fluorescence images of transgenic zMPO:GFP embryos at various stages of development are shown, with anterior/head pointing left and dorsal side facing right (A, B) or upward (C, D). (A) Embryo at the 9-somite stage; arrow indicates signal seen in the anterior lateral mesoderm, arrowhead indicates signal seen in the tailbud region. (B) Composite image of an embryo at the 19-somite stage (lateral view, dorsal side to the left); in the upper half arrows indicate GFP+ cells over the yolk, in the lower half GFP+ cells in the Intermediate Cell Mass (ICM) are indicated. Arrowhead indicates GFP expression in the tail. (C) Embryo at 48 hpf, arrowhead indicates the heart. (D) Higher magnification of area boxed in (C), detailing the ICM and tail. Embryo in C-D was maintained in media containing 0.2 mM PTU to prevent pigment formation. Note that the GFP expression in cells around the head, yolk sac and ICM are consistent with previous observations of zMPO mRNA 8, 9; the GFP expression seen during somitogenesis (A), in the developing tail (B) and in the heart and tailfin cells (C, D) do not correspond with these reports.
• Supplemental Movie 1. - Supplemental Movie 1. Wild-type leukocyte response to fin wound. A wild-type zebrafish embryo at 3 dpf was wounded in the ventral fin and observed by time-lapse DIC microscopy; the region imaged is the ventral fin in the midsection of the embryo, just below the vasculature and ICM. Note the migration of apparent leukocytes from the vasculature/ICM to the wound site.
• Supplemental Movie 2. - Supplemental Movie 2. zMPO:GFP cells within the Intermediate Cell Mass. The midbody of a zMPO:GFP embryo at 3 dpf is shown, anterior to the left and dorsal side pointing up. The movie was made at 10 s per frame over 20 min. The first image is an Oblique Coherent Contrast image overlaid with fluorescence; all images thereafter are only fluorescence. Note the movement of GFP+ cells within the Intermediate Cell Mass, and the presence of these cells within the circulation of the embryo.
• Supplemental Movie 3. - Supplemental Movie 3. zMPO:GFP cell response to a wound. The ventral fin of a zMPO:GFP embryo at 3 dpf is shown, anterior to the left and dorsal side pointing up. The movie was made at 1 min per frame over 4 h (240 frames total), starting 5 min after a wound was made in the fin. Differential Interference Contrast and fluorescent images were captured as described in materials and methods. Note the migration of GFP+ cells both toward and away from the wound, and within the ICM. Also note the GFP-expressing fin cell to the left of the wound, which does not migrate during the movie.