DEV01667 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Schematic diagram showing how the position of an exemplary cell within the germ ring was determined. The cell movement tracks within the embryo (blue tracks) were assembled using the x1 and y1 coordinates, while the tracks for the movements of cells within the germ ring (red tracks) were assembled by calculating the changes in x3 (x2-x1) and y3 (y2-y1) over time with x1,y1 as starting point.

• Supplemental Figure 2 -

  Fig. S2. Summary of the movement tracks of all cells analyzed in supplementary movies. Panels A-D correspond to supplementary movies 1-4, respectively. In A-D, the blue tracks delineate the movement of epiblast cells and the red tracks the movement of hypoblast cells within the embryo (corresponding to blue tracks in Figs 2, 5). In A, cells closer to the surface of the blastoderm were labeled as ‘epiblast’ (red) and cells closer to the yolk cell as ‘hypoblast’ (blue), although the blastoderm at this stage cannot yet formally be divided into epiblast and hypoblast. The yellow dots indicate the starting points of the cell tracks. Cells were tracked in 2.6-minute time intervals. Please note that the blue/red coding of cell tracks in this figure is different from the one in Figs 2 and 5.

• Supplemental Figure 3 -

  Fig. S3. Western blot showing the total amounts of E-Cadherin protein from wild-type and e-cadherin morpholino (MO; 8ng)-injected embryos at bud (10 hpf) and shield (6 hpf) stages. The size of the band recognized corresponds to the predicted molecular weight for E-Cadherin (140 kDa) (Babb and Marrs, 2004). wt, wild type; MO, e-cadherin MO. Two independent samples for each condition with 10 embryos/sample were loaded. As a loading control, the gels were Commassie stained to check for equal amounts of protein in each lane.

• Movie 1 -

  Movie 1. Time-lapse movie of the cell movements within the germ ring margin at the region of the shield from the onset of gastrulation (40% epiboly) onwards. The movie was recorded for a total of 68 minutes in 2.6-minute time intervals. Sagittal views. Frame size=2063206 mm.

• Movie 2 -

  Movie 2. Time-lapse movie of the cell movements within the germ ring margin at the region of the shield from shield stage (60% epiboly) onwards. The movie was recorded for a total of 123 minutes in 2.6-minute time intervals. Sagittal views. Frame size=2063206 mm.

• Movie 3 -

  Movie 3. Time-lapse movie of the cell movements within the germ ring margin of an e-cadherin MO-injected embryo at the region of the shield from early shield stage (6 hpf) onwards. The movie was recorded for a total of 105 minutes in 2.6-minute time intervals. Sagittal views. Frame size=2063206 mm.

• Movie 4 -

  Movie 4. Time-lapse movie of the cell movements within the germ ring margin of an e-cadherin MO-injected embryo at the region of the shield from late shield stage (7.5 hpf) onwards. The movie was recorded for a total of 63 minutes in 2.6-minute time intervals. Sagittal views. Frame size=2063206 mm.

• Movie 5 -

  Movie 5. Time-lapse movie showing the aggregation behavior of cultured primary mesendodermal (cyc-overexpressing) cells from sphere stage wild-type (red) and e-cadherin MO-injected (green) embryos during the first 7.5 hours in culture.

• Movie 6 -

  Movie 6. Time-lapse movie showing the aggregation behavior of cultured primary ectodermal (mz-oep) cells from sphere stage wild-type (red) and e-cadherin MO-injected (green) embryos during the first 7.3 hours in culture.