DEV020107 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. The endodermal cells lie in the deepest layer of the hypoblast. (A) Confocal image of endodermal and mesodermal cells in Tg(sox17:EGFP) embryo at the 90% epiboly stage. Dextran-Alexa594 was injected into a Tg(sox17:EGFP) embryo at the one-cell stage. The endodermal cells, which have many filopodial processes, are yellowish green and mesodermal cells are red. (B) Reconstructed z-section at the level of the dashed line in A. The extra-embryonic YSL is also red. The endodermal cells occupy the space between the mesodermal cells and extra-embryonic YSL. (C) Schematic representation of B. YSL, yolk syncytial layer.

• Supplemental Figure S2 -

  Fig. S2. Sdf1a and Sdf1b act redundantly on endodermal cell migration. (A-D) Dorsal views of the mid-trunk region of Tg(sox17:EGFP) transgenic embryos at the three-somite stage. Anterior is to the top. The delay of endodermal dorsal migration was more severe in sdf1a/sdf1b double-knockdown embryos than in single-knockdown embryos of either sdf1a or sdf1b. Moreover, the phenotype on the dorsal migration of endodermal cells in sdf1a knockdown embryos is a little milder than that of sdf1b knockdown embryos.

• Supplemental Figure S3 -

  Fig. S3. Phosphoinositide 3-kinase (PI3K) signaling does not regulate the movements of endodermal cells. (A-F) Confocal images of endodermal cells in Tg(sox17:EGFP) transgenic embryos at the three-somite stage. Dorsal views, anterior to the top. One marginal blastomere cell at the 128-cell stage has been transformed into an endodermal fate by the misexpression of cas mRNA (2 pg). The injected cell was labeled with dextran-Alexa594. (A-C) The progeny of the injected cell intermingle with EGFP-expressing endodermal cells. (D-F) The migration of cells co-injected with both cas (2 pg) and a dominant-negative form of phosphoinositide 3-kinase (dnPI3K) (4 pg) mRNAs was not changed relative to cells injected with cas mRNA alone.

• Movie 1 -

  Movie 1. Time-lapse movie of migrating endodermal cells in a Tg(sox17:EGFP) transgenic embryo from 65% to 90% epiboly stages. Lateral views with dorsal to the right.

• Movie 2 -

  Movie 2. Time-lapse movie of migrating endodermal cells in a Tg(sox17:EGFP) transgenic embryo from 90% epiboly to tail bud (TB) stages. Lateral views with dorsal to the right.

• Movie 3 -

  Movie 3. Time-lapse movie of migrating endodermal cells in a Tg(sox17:EGFP) transgenic embryo from 65% epiboly to TB stages. The transplanted nlacZ-expressing cells are red due to the lineage tracer dextran-Alexa594. The nlacZ-expressing cells do not attract the endodermal cells. AP, animal pole; VP, vegetal pole; V, ventral; D, dorsal. Scale bar: 100 µm.

• Movie 4 -

  Movie 4. Time-lapse movie of migrating endodermal cells in a Tg(sox17:EGFP) transgenic embryo from 65% epiboly to TB stages. The transplanted sdf1a-expressing cells are red due to the lineage tracer dextran-Alexa594. The endodermal cells are attracted towards the sdf1a-expressing cells. AP, animal pole; VP, vegetal pole; V, ventral; D, dorsal. Scale bar: 100 µm.

• Movie 5 -

  Movie 5. Time-lapse movie of migrating endodermal cells in a Tg(sox17:EGFP) transgenic embryo from 65% epiboly to TB stages. The transplanted sdf1b-expressing cells are red due to the lineage tracer dextran-Alexa594. The endodermal cells are attracted towards the sdf1b-expressing cells. AP, animal pole; VP, vegetal pole; V, ventral; D, dorsal. Scale bar: 100 µm.

• Movie 6 -

  Movie 6. Time-lapse movie of migrating endodermal cells in a misMO1-injected Tg(sox17:EGFP) transgenic embryo at 90% epiboly stage. The endodermal cells in the lateral region of these morphants were analyzed. The direction of migration is to the right. Scale bar: 20 µm.

• Movie 7 -

  Movie 7. Time-lapse movie of migrating endodermal cells in a cxcr4aMO1-injected Tg(sox17:EGFP) transgenic embryo at 90% epiboly stage. The endodermal cells in the lateral region of these morphants were analyzed. The direction of migration is to the right. Scale bar: 20 µm.

• Movie 8 -

  Movie 8. Time-lapse movie of migrating endodermal cells in a sdf1MOs-injected Tg(sox17:EGFP) transgenic embryo at 90% epiboly stage. The endodermal cells in the lateral region of these morphants were analyzed. The direction of migration is to the right. Scale bar: 20 µm.