DEV02849 Supplementary Material

Files in this Data Supplement:

• Supplemental Tables 1-2 - Adobe PDF

• Supplemental Figure S1 -

  Fig. S1. Effects of Ena inactivation on early development and imaginal discs. Anterior to left. Antigens as indicated. (A-L) Early development in mat-FP4mito or ena²³ maternal mutants. (A,B) Wild-type cellularization. (C,D) ena²³ maternal mutant. (E-I′) mat-FP4mito. (E,F) Cellularizing embryo with few defects. (G,H) Syncytial embryo with elevated nuclear loss (small actin rings without associated nuclei; arrows). (I) Embryo with uneven nuclear distribution at anterior end (arrow). (J) Ena, wild-type cellularization. Ena is largely diffusely cytoplasmic, with occasional slight enrichment apically and at the cellularization front. (K) mat-FP4mito, cellularization. Ena recruited by FP4mito apical to nuclei (arrowhead). (L) ena²³ maternal mutant. (M-O) Ena inactivation does not disrupt wing imaginal disc epithelia. FP4mito expressed in posterior compartment with engrailed-Gal4. (M) Wild-type Ena at AJs (arrows). FP4mito recruitment to presumptive mitochondria (arrowheads). (N) Surface view. (O) Cross-section, epithelial fold. Arrows, apical AJs/actin; arrowheads, boundary, FP4mito expression. Scale bars: 20 μm in A-L; 10 μm in M-O.

• Supplemental Figure S2 -

  Fig. S2. GFP-Ena in retracting filopodia. Movie stills showing an embryo expressing both GFP-Ena and RFP-actin using engrailed-Gal4. Arrowhead, GFP-Ena at filopodial tip; arrow, GFP-Ena particles moving backward from tip. Time is in minutes and seconds. Scale bar: 20 μm.

• Movie 1 -

  Movie 1. High-magnification view, dorsal closure-staged embryo expressing GFP-Ena using engrailed-Gal4. Spots of GFP-Ena move away from the tip of the filopodium just before it starts to retract. A subset of the GFP-Ena remains at the tip of the filopodium as it retracts. Images collected every 5 seconds; Movie displayed at 10 frames/second. Time is in minutes and seconds. Still images from this Movie are shown in Fig. 7B.

• Movie 2 -

  Movie 2. High-magnification view, dorsal closure-staged embryo expressing GFP-actin+GFP-Ena. A large fan-like lamellipodium containing numerous actin microspikes is produced by leading-edge cells. As this structure resolves, several microspikes fuse at their distal tips to form filopodia. Images collected every 15 seconds; Movie displayed at 10 frames/second. Time is in minutes and seconds. Still images from this Movie are shown in Fig. 7C.

• Movie 3 -

  Movie 3. Dorsal closure-staged embryo expressing GFP-actin using e22c-Gal4. Leading edges of wild-type migrating epithelial sheets as they meet and fuse at the dorsal midline. Images collected every 15 seconds; Movie displayed at 10 frames/second. Still images from this Movie are shown in Fig. 8A.

• Movie 4 -

  Movie 4. Dorsal closure-staged embryo expressing GFP-actin+FP4mito using e22c-Gal4. Although leading edges of the migrating epithelial sheets eventually meet and fuse at the dorsal midline, they have difficulties zippering. Images collected every 15 seconds; Movie displayed at 10 frames/second. Still images from this Movie are shown in Fig. 8B.

• Movie 5 -

  Movie 5. High-magnification view, dorsal closure-staged embryo expressing GFP-actin using engrailed-Gal4. Leading-edge cells in wild-type embryos produce broad lamellipodia from which thin filopodia are produced. Images collected every 15 seconds; Movie displayed at 10 frames/second. Time is in minutes and seconds. Still images from this Movie are shown in Fig. 9A.

• Movie 6 -

  Movie 6. High-magnification view, dorsal closure-staged embryo expressing GFP-actin+FP4mito using engrailed-Gal4. Leading-edge cells produce predominantly lamellipodia and a small number of short filopodia. Images collected every 15 seconds; Movie displayed at 10 frames/second. Time is in minutes and seconds. Still images from this Movie are shown in Fig. 9B.

• Movie 7 -

  Movie 7. High-magnification view, dorsal closure-staged embryo expressing GFP-actin+UAS-Ena using engrailed-Gal4. Leading-edge cells overexpressing Ena produce longer filopodia than wild type. Images collected every 15 seconds; Movie displayed at 10 frames/second. Time is in minutes and seconds. Still images from this Movie are shown in Fig. 9D.