Supplementary Materials

Tissue topography steers migrating Drosophila border cells

Wei Dai, Xiaoran Guo, Yuansheng Cao, James A. Mondo, Joseph P. Campanale, Brandon J. Montell, Haley Burrous, Sebastian Streichan, Nir Gov, Wouter-Jan Rappel, Denise J. Montell

Materials/Methods, Supplementary Text, Tables, Figures, and/or References

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  • Materials and Methods
  • Supplementary Text ST1 to ST3
  • Figs. S1 to S23
  • Tables S1 to S11
  • Captions for Movies S1 to S10
  • References

Images, Video, and Other Media

Movie S1
Confocal time-lapse imaging of normal border cell migration. SlboGal4 drives UAS-mCD8-EGFP (green) and UAS-RFPnls (magenta).
Movie S2
"Fly-through" from the anterior tip of the egg chamber where the border cells are located to the oocyte border, using near isotropic light sheet imaging showing the nurse cell membranes encountered by the border cells as they migrate. Multiple nurse cells meet in the center of the egg chamber where there is high membrane curvature. Border cells encounter ~40 side paths along the way. Green, E-cadherin. Magenta, F-actin. Scale bar, 20 μ m.
Movie S3
Confocal time-lapse imaging of border cell migration with germline knockdown of E-cadherin. Matalpha4-Gal4;UAS-EcadRNAi. Green, Slbo-PHEGFP; magenta, UMAT-Lyn-tdTomato.
Movie S4
(Fig. 2E and F) (A) Confocal time-lapse imaging of a nurse cell juncture pulled by an incoming border cell cluster in a control egg chamber. Green, Slbo-PHEGFP; magenta, UMAT-Lyn-tdTomato, Ubi-HisRFP. Note that the juncture shown by the yellow filament is just an example. Other junctures that the border cells contact are also deflected. In the first and last time points, tracking is removed to show the original membrane signal. (B) Confocal time-lapse imaging of an egg chamber with germline knockdown of E-cadherin (Matalpha4-Gal4;UAS-EcadRNAi) showing absence of nurse cell membrane deflection when border cell protrusions touch nurse cell membranes. Despite touching the nurse cell juncture multiple times, the border cell protrusion did not deflect it (one juncture is highlighted by the yellow filament). In the first and last time points, tracking is removed to show the original membrane signal.
Movie S5
(fig. S11B, fig. S19A) Animation of 3-dimensional reconstruction of nurse cells and ring canals in a stage 9 egg chamber. White, F-actin; green, Slbo-LifeactGFP; blue, Hoechst. Nurse cell color indicates connection distance to the oocyte (red). White, border cells at the anterior tip. Yellow circles, ring canals. The egg chamber was stained to show F-actin (white), border cells (green), and nuclei (blue). First the reconstructed nurse cells rotate 360 degrees to show that nurse cells that connect closer to the oocyte (red) are located more posteriorly. Then the ring canals (yellow) are shown to display their location in between connected nurse cells.
Movie S6
(Fig. 3A and C) Animation of 3-dimensional reconstruction of different types of nurse cell contacts in a stage 9 egg chamber. Green, Slbo-LifeactGFP; white, F-actin or F-actin with outer follicle cell signal masked. Two-cell-contacts are shown as surfaces and color represents position along the z axis. Three-nurse cell junctures are represented as lines and color provides position in z. Magenta dots represent junctures of >3 nurse cells. First F-actin (white) and border cells (green) are shown. Then outer-follicle cell F-actin signals are masked to show nurse cell contacts. Then 2-cell surfaces are shown in 360 degree rotation, followed by >=3 cell juncture views in another 360 degree rotation.
Movie S7
(Fig. 4A) Simulation of border cell migration with nurse cell geometry and anterior-posterior chemoattractant gradient.
Movie S8
(fig. S17B and C). A confocal time-lapse movie of border cell migration in germline knockdown of E-cadherin. Border cells migrate along nurse-cell/nurse-cell/follicle-cell paths instead of nurse-cell/follicle-cell. Green, Slbo-PHEGFP; magenta, dextran.
Movie S9
(fig.S22A) A confocal time-lapse movie of wild-type border cell migration. Note the dorsal migration prior to reaching the oocyte boundary. Green, Slbo-PHEGFP; white, UMAT-Lyn-tdTomato.
Movie S10
(fig.S22C) Simulation of border cell migration with nurse cell geometry and anterior-posterior chemoattractant gradient and dorsal Grk gradient.