Polarized E-cadherin endocytosis directs actomyosin remodeling during embryonic wound repair

Files in this Data Supplement:

• Supplemental Material (PDF)
• Video 6 -
  Delayed wound closure in shi^ts1 embryos heated to the restrictive temperature. Epidermal cells expressing GFP:moesin in a wild-type embryo heated to the restrictive temperature (35°C, left), a shi^ts1 embryo imaged at the permissive temperature (23°C; middle), or a shi^ts1 embryo heated to the restrictive temperature (35°C; right). Images were acquired by time-lapse confocal microscopy using a spinning disk confocal microscope (Revolution XD; Andor Technology). A stack was acquired every 15 s for 42.5 min. Time after wounding is shown. Anterior left, dorsal up.
• Video 7 -
  BAPTA injection prevents extracellular calcium release upon wounding. Epidermal cells expressing GCaMP3, a fluorescent calcium biosensor, in embryos injected with water (left) or 50 mM BAPTA (right). Images were acquired by time-lapse confocal microscopy using a spinning disk confocal microscope (Revolution XD; Andor Technology). A stack was acquired every 30 s for 41.5 min. Time after wounding is shown. Anterior left, dorsal up.
• Video 8 -
  Calcium is required for polarized accumulation of dynamin at the wound margin. Epidermal cells in embryos expressing dynamin:GFP, injected with water (left) or 50 mM BAPTA (right). Images were acquired by time-lapse confocal microscopy using a spinning disk confocal microscope (Revolution XD; Andor Technology). A stack was acquired every 30 s for 51.5 min. Time after wounding is shown. Anterior left, dorsal up.
• Video 1 -
  Myosin accumulates at the wound margin. Epidermal cells expressing myosin:GFP. Images were acquired by time-lapse confocal microscopy using a spinning disk confocal microscope (Revolution XD; Andor Technology). A stack was acquired every 15 s for 41 min. Time after wounding is shown. Anterior left, dorsal up.
• Video 9 -
  Calcium is required for E-cadherin removal from the wound margin. Epidermal cells in embryos expressing endo–E-cadherin:GFP, injected with water (left) or 50 mM BAPTA (right). Images were acquired by time-lapse confocal microscopy using a spinning disk confocal microscope (Revolution XD; Andor Technology). A stack was acquired every 30 s for 42 min. Time after wounding is shown. Anterior left, dorsal up.
• Video 2 -
  Clathrin accumulates at the wound margin. Epidermal cells expressing GFP:clc. Images were acquired by time-lapse confocal microscopy using a spinning disk confocal microscope (Revolution XD; Andor Technology). A stack was acquired every 30 s for 41.5 min. Time after wounding is shown. Anterior left, dorsal up.
• Video 4 -
  ARF6 accumulates at the wound margin. Epidermal cells expressing ARF6:GFP. Images were acquired by time-lapse confocal microscopy using a spinning disk confocal microscope (Revolution XD; Andor Technology). A stack was acquired every 30 s for 41.5 min. Time after wounding is shown. Anterior left, dorsal up.
• Video 5 -
  Dynasore treatment impairs wound closure. Epidermal cells expressing ubi–E-cadherin:GFP (green) and myosin:mCherry (red) in a control embryo injected with 50% DMSO (left) or 50 mM dynasore (right). Images were acquired by time-lapse confocal microscopy using a spinning disk confocal microscope (Revolution XD; Andor Technology). A stack was acquired every 30 s for 44 min. Time after wounding is shown. Anterior left, dorsal up.
• Video 3 -
  Dynamin accumulates at the wound margin. Epidermal cells expressing dynamin:GFP. Images were acquired by time-lapse confocal microscopy using a spinning disk confocal microscope (Revolution XD; Andor Technology). A stack was acquired every 30 s for 53.5 min. Time after wounding is shown. Anterior left, dorsal up.