Supplementary Materials

Adhesion Functions in Cell Sorting by Mechanically Coupling the Cortices of Adhering Cells

Jean-Léon Maître, Hélène Berthoumieux, Simon Frederik Gabriel Krens, Guillaume Salbreux, Frank Jülicher, Ewa Paluch, Carl-Philipp Heisenberg

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

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  • Supplementary Text
  • Materials and Methods
  • Figs. S1 to S10
  • Legends for movies S1 to S23
  • Tables S1 to S12
  • References

Images, Video, and Other Other Media

Movie S1
Homotypic ectoderm cell doublet during the first 10 min of contact formation. Bright-field images are recorded with 1 s time-intervals and displayed at 60 frames per s (fps). Scale bar = 10 μm.
Movie S2
Homotypic mesoderm cell doublet during the first 10 min of contact formation. Bright-field images are recorded with 1 s time-intervals and displayed at 60 frames per s (fps). Scale bar = 10 μm.
Movie S3
Homotypic endoderm cell doublet during the first 10 min of contact formation. Bright-field images are recorded with 1 s time-intervals and displayed at 60 frames per s (fps). Scale bar = 10 μm.
Movie S4
Homotypic ectoderm triplet during cell-cell separation, showing the change in curvature at the former cell-cell contact shortly after separation. Bright-field images are recorded with 1 s time-intervals and displayed at 10 frames per s (fps). Scale bar = 10 μm.
Movie S5
Homotypic mesoderm triplet during cell-cell separation, showing the change in curvature at the former cell-cell contact shortly after separation. Bright-field images are recorded with 1 s time-intervals and displayed at 10 frames per s (fps). Scale bar = 10 μm.
Movie S6
Homotypic endoderm triplet during cell-cell separation, showing the change in curvature at the former cell-cell contact shortly after separation. Bright-field images are recorded with 1 s time-intervals and displayed at 10 frames per s (fps). Scale bar = 10 μm.
Movie S7
Separation force measurement of a homotypic ectoderm doublet. Bright-field images are recorded with 1 s time-interval and displayed at 10 frames per s (fps). Scale bar = 10 μm.
Movie S8
Separation of a Cdh2-eGFP expressing ectoderm doublet. Cdh2-eGFP accumulates at the cell-cell contact during separation. Once the cell-cell contact is dissolved, high levels of Cdh2-eGFP are found at the base of plasma membrane-tethers connecting the separated cells. Confocal images are recorded at 580 ms time‐intervals, averaged over a total period of 5.2 s and displayed at 2 frames per s (fps). Scale bar = 10 μm.
Movie S9
Separation of an eGFP-Ctnnb1 expressing ectoderm doublet. eGFP-Ctnnb1 accumulates at the cell-cell contact during separation. Once the cell-cell contact is dissolved, high levels of eGFP-Ctnnb1 are found at the base of plasma membrane-tethers connecting the separated cells. Confocal images are recorded at 580 ms time‐intervals, averaged over a total period of 5.2 s and displayed at 2 frames per s (fps). Scale bar = 10 μm.
Movie S10
Separation of a Ctnna-eGFP expressing ectoderm doublet. Ctnna-eGFP does not accumulate at the cell-cell contact during separation. Once the cell-cell contact is dissolved, high levels of Ctnna-eGFP are found at the base of plasma membrane-tethers connecting the separated cells. Confocal images are recorded at 580 ms time‐intervals, averaged over a total period of 5.2 s and displayed at 2 frames per s (fps). Scale bar = 10 μm.
Movie S11
Separation of a LifeAct-RFP expressing ectoderm doublet. LifeAct-RFP does not accumulate at the cell-cell contact during separation. Once the cell-cell contact is dissolved, high levels of LifeAct-RFP are found at the base of plasma membrane-tethers connecting the separated cells. Confocal images are recorded at 580 ms time‐intervals, averaged over a total period of 5.2 s and displayed at 2 frames per s (fps). Scale bar = 10 μm.
Movie S12
Cdh2-eGFP expressing ectoderm doublets lacking endogenous Cdh1 expression (cdh1 morphant cells) during the first 5 min of contact formation. Confocal images are recorded at 290 ms time-intervals, averaged over a total period of 5.2 s and displayed at 10 frames per s (fps). Scale bar = 10 μm.
Movie S13
Cdh2-eGFP expressing endoderm doublets lacking endogenous Cdh1 expression (cdh1 morphant cells) during the first 5 min of contact formation. Confocal images are recorded at 290 ms time-intervals, averaged over a total period of 5.2 s and displayed at 10 frames per s (fps). Scale bar = 10 μm.
Movie S14
Cdh2Δcyto-eGFP expressing ectoderm doublets lacking endogenous Cdh1 expression (cdh1 morphant cells) during the first 5 min of contact formation. Confocal images are recorded at 290 ms time-intervals, averaged over a total period of 5.2 s and displayed at 10 frames per s (fps). Scale bar = 10 μm.
Movie S15
Cdh2Δcyto-eGFP expressing endoderm doublets lacking endogenous Cdh1 expression (cdh1 morphant cells) during the first 5 min of contact formation. Confocal images are recorded at 290 ms time-intervals, averaged over a total period of 5.2 s and displayed at 10 frames per s (fps). Scale bar = 10 μm.
Movie S16
Sorting of Cdh2-eGFP expressing ectoderm labeled with cytoplasmic Rhodamin‐ or Alexa627-Dextrans. Homotypic cell sorting of differently labeled (displayed in red and green) Cdh2-eGFP expressing ectoderm cells lacking endogenous Cdh1 (cdh1 morphant). ~ 4 μm spaced confocal slices are recorded every ~ 5 min for 5 h, and displayed as a 3D render at 2 frames per s (fps). Scale bar = 10 μm.
Movie S17
Sorting of Cdh2-eGFP or Cdh2Δcyto-eGFP expressing ectoderm labeled with cytoplasmic Rhodamin- or Alexa627-Dextrans. Heterotypic cell sorting of of Cdh2-eGFP (displayed in red and green) or Cdh2Δcyto-eGFP expressing ectoderm cells lacking endogenous Cdh1 (cdh1 morphant). ~ 4 μm spaced confocal slices are recorded every ~ 5 min for 5 h, and displayed as a 3D render at 2 frames per s (fps). Scale bar = 10 μm.
Movie S18
Sorting of Cdh2-eGFP expressing endoderm labeled with cytoplasmic Rhodamin‐ or Alexa627-Dextrans. Homotypic cell sorting of differently labeled (displayed in red and green) Cdh2-eGFP expressing ectoderm cells lacking endogenous Cdh1 (cdh1 morphant). ~ 4 μm spaced confocal slices are recorded every ~ 5 min for 5 h, and displayed as a 3D render at 2 frames per s (fps). Scale bar = 10 μm.
Movie S19
Sorting of Cdh2-eGFP or Cdh2Δcyto-eGFP expressing endoderm labeled with cytoplasmic Rhodamin‐ or Alexa627-Dextrans. Heterotypic cell sorting of of Cdh2-eGFP (displayed in red and green) or Cdh2Δcyto-eGFP expressing ectoderm cells lacking endogenous Cdh1 (cdh1 morphant). ~ 4 μm spaced confocal slices are recorded every ~ 5 min for 5 h, and displayed as a 3D render at 2 frames per s (fps). Scale bar = 10 μm.
Movie S20
Sagittal view of the shield region in a Myl12.1-eGFP expressing transgenic embryo at 6 hpf. Myl12.1-eGFP accumulates at the interface between epiblast and hypoblast. Increased levels of Myl12.1-eGFP can also be found at the cleavage furrow of dividing cells, the EVL/YSL margin, and at retracting blebs. In contrast, Myl12.1-eGFP localization is reduced at cell-cell contacts. Two-photon images are recorded at 30 s time-intervals and displayed at 10 frames per s (fps). Scale bar = 10 μm.
Movie S21
3D rendering (Imaris, Bitplane) of a confocal image stack of epiblast cells at the animal pole of an embryo at 5 hpf (ectoderm) stained 40 with α-Ctnna antibodies. Note the ring-like distribution of Ctnna at the margin of cell-cell contacts. Scale-bar = 20 μm.
Movie S22
Cells within the lateral mesendoderm of a transgenic embryo expressing hRas-eGFP at 6 hpf. Note the plasma membrane tethers forming between separating cells. Two-photon images at 1 min time-intervals and displayed at 2 frames per s (fps). Scale bar = 10 μm.
Movie S23
Epiblast cells at the animal pole of an embryo expressing Cdh2-eGFP. Note Cdh2-eGFP accumulation at plasma membrane tethers between separating cells. Two-photon images at 13 s time-intervals and displayed at 5 frames per s (fps). Scale bar = 10 μm.