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. 2018 Mar 21;8:4986. doi: 10.1038/s41598-018-22940-3

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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E-cadherin facilitates infiltration of 4T1 cell into stromal cells, ex vivo. GFP-expressing 4T1 cells (Con) or shEcad 4T1 cells (shEcad), and RFP expressing mouse embryo fibroblasts (MEF) were plated in two parallel compartments of a silicon insert (Ibidi®). When the cells within the two compartments reached confluence, the insert was removed, and the area between the two cell monolayers was imaged by live-cell video microscopy. (A) Snapshots taken at the indicated time points, showing the Con (left column, yellow) and shEcad (right column, yellow) and the MEF cells (magenta-colored). The vertical wavy line marks the position of the cancer cells at t = 0. Scale bar: 100 μm. (B) Plot showing the positions of the “invasive front” of the shEcad (blue) and Con (red) over a period of 65 hours. The times at which the shEcad and Con 4T1 cells physically reach the fibroblasts are indicated by the blue and red arrows, respectively. Note that the initial migration rate of the shEcad cells is considerably faster than that of the control cells, while their infiltration into the stromal monolayer is considerably slower than that of the E-cadherin-expressing and tether-forming Con cells. For the full movie, see Supplementary Movie S10.