DEV030866 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 - Adobe PDF

• Supplemental Figure S1 -

  Fig. S1. Dynamics of AS contraction in individual wild-type and ASGal4 embryos. Each embryo is represented by a different coloured ribbon, representing the mean±s.e. (A,B) Wild type; (C,D) ASGal4/UAS-spastin; (E,F) ASGal4/UAS-p35. (A,C,E) Apical cell area; (B,D,F) proportional rate of area change. Data pooled for each genotype are presented in Fig. 1.

• Supplemental Figure S2 -

  Fig. S2. Bazooka localization in AS cells. Stage 13 (A-A′′) and stage 14 (B-B′′) embryos stained with antibodies against Baz (green, shown as a single channel in A′′,B′′) and Ecad (red).

• Supplemental Figure S3 -

  Fig. S3. Apical contraction in ML and AP directions. (A) Proportional rates of size change of AS cells in ML (red) and AP (blue) orientations for data pooled from three ASGal4/UAS-p35 embryos. (B,C) Proportional rates of size change in ML (B) and AP (C) orientation for data pooled from four wild-type (orange ribbons) and three ASGal4/UAS-p35 (magenta ribbons) embryos.

• Supplemental Figure S4 -

  Fig. S4. Differences in cell behaviour along the AP axis in ASGal4/UAS-p35 embryos. (A-I) Panels are the same as in Fig. 4, but here for ASGal4/UAS-p35 embryos for the time ranges shown.

• Supplemental Figure S5 -

  Fig. S5. Defects in the attachment of the AS to the yolk when a dominant-negative adhesive form of βPSintegrin is ectopically expressed in the yolk. Thick transverse sections of a wild-type (left) and a P0180/UAS-TorsoβPScyt (right) embryo stained with antibodies against Ecad and Scribbled. The dashed line indicates the limit of the yolk.

• Supplemental Figure S6 -

  Fig. S6. Dynamics of AS contraction in mys mutant embryos. (A,B) Apical cell area (A) and proportional rate of area change (B). Each embryo is represented by a different coloured ribbon, representing the mean±s.e. Pooled data for each genotype are presented in Fig. 7. (C-K) Differences in cell behaviour along the AP axis in mys embryos. (C) Proportional change in area/minute (area strain rate) of AS cells as a function of their AP location over time. Data are pooled from four aligned mys embryos. (D,E) Mean proportional rates of change in the ML orientation for cells across the AS for the periods 30-60 minutes and 90-120 minutes after the start of dorsal closure, respectively. Data are pooled from four aligned mys embryos. Colours represent the mean behaviour of cells that fall within each tile of AS tissue. (D′,E′) The same data as in D,E are presented as averages over the ML (red) and AP (light blue) axis, for the same periods of DC. (F-H) Still images from an animation of an example mys embryo, showing the relative magnitude and orientation of the long axis of AS cells. (I-K) Cell orientation data are pooled from four aligned mys embryos, and regional averages are shown for three epochs of DC, 0-30, 60-90 and 120-150 minutes after the onset of DC, respectively. The orientation of the red lines represents the mean (elongation ratio-weighted) orientation of the long axis of cells in each grid square. The lengths of the red lines represent the mean elongation log-ratio of the long-to-short axes of cell shapes for cells within each grid square. A line length equal to the size of a grid square equals a log-ratio of 1.0 (a ratio of 2.718:1).

• Supplemental Figure S7 -

  Fig. S7. Cell extrusion contributes to the radial pattern of AS cell apical contraction in mys mutant embryos. (A,B) The same graphs as in Fig. 7E,F for four aligned mys embryos, but cells that extrude from the AS (e.g. gold cells in Fig. 4A) have been removed from the analysis. An inverse gradient is still observed in B.

• Movie 1 -

  Movie 1. Time-lapse of a wild-type embryo carrying a ubiECadGFP transgene during dorsal closure. Time interval between frames in this and all other movies: 2 minutes.

• Movie 2 -

  Movie 2. Time-lapse of an enGal4/UAS-spastin-EGFP embryo carrying a ubiECadGFP transgene during dorsal closure.

• Movie 3 -

  Movie 3. Time-lapse of an ASGal4/UAS-p35 embryo carrying a ubiECadGFP transgene during dorsal closure.

• Movie 4 -

  Movie 4. Time-lapse of a mys mutant embryo carrying a ubiECadGFP transgene during dorsal closure.

• Movie 5 -

  Movie 5. Time-lapse of a mys mutant embryo carrying a UASactinGFP transgene under the control of the enGal4 line during dorsal closure.

• Movie 6 -

  Movie 6. Time-lapse of a P0180/UAS-TorsoβPScyt embryo carrying a ubiECadGFP transgene during dorsal closure.