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. 2022 Oct 28;13:6423. doi: 10.1038/s41467-022-34165-0

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© The Author(s) 2022

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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Fig. 2 — a–d Filopodia interact with epithelial vertices as MCC moves into the superficial epithelium. The actin cortex of MCCs is labeled with α-tubulin::LifeAct-RFP (pseudo-colored in green in composite images and gray as a separate channel). a Image sequence of integrating MCC interacting with overlying vertices. Epithelial vertices are labeled with lipolysis stimulated lipoprotein receptor tagged with x3GFP (LSR, pseudo-colored in magenta in composite images and fire as a separate channel). Scale bar: 10 μm. White dotted lines outline overlying junctions. The yellow arrowheads depict the orientation used for 3D rendering in b. b 3D rendering of a, with MCC forming contacts with different vertices (marked by white arrows). White lines outline overlying junctions. c Orthogonal (XZ) projections of a, depicting the attachment between filopodia (marked by white arrows) and vertices. Yellow boxes mark insets for separate channels. Scale bar: 5 μm. d Close-up of LSR-GFP (pseudo-colored in magenta in composite and fire as a separate channel) localization within a growing and retracting filopodium, visualized by F-actin marker (LifeAct, pseudo-colored in green). LSR is visualized by expressing α-tubulin::LSR-GFP. Scale bar: 2 μm. e Orthogonal (XZ) projections of filopodium pulling on the epithelial vertex (marked by white arrows). The epithelial vertex is marked by expressing nectin::LSR-GFP. The white dotted line outlines the MCC contour and the yellow dotted line outlines the apical surface of the superficial epithelium. Scale bar: 2 μm. f Quantification of vertex pulling from e. The MCC F-actin intensity (green) and vertex length during one event of vertex pulling (purple) and for a non-pulled vertex (magenta). g Schematics representing MCC probing and vertex pulling.