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. 2019 May 9;10:2115. doi: 10.1038/s41467-019-09676-y

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© The Author(s) 2019

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. 6 — KRT80 induces invasion-associated cytoskeletal changes. a Representative confocal microscopy images showing F-actin (magenta), KRT80 (green) and DAPI (blue) staining of MCF7-control, MCF7-K80, LTED-control and LTED-sh^a cells. Scale bars represent 25 μm. b Zoom-up magnifications of areas indicated in a, showing F-actin (magenta), KRT80 (green) and DAPI (blue) staining in cells located at the border of clusters. Single channel images for F-actin and KRT80 are also shown. Scale bars, 10 μm. Asterisks indicate lamellipodia-like structures in MCF7-K80 and LTED cells, and hashtags indicate cortical actin areas in MCF7 and LTED-sh^a cells. Graphs on the right show line scan analysis for F-actin and KRT80 fluorescence across the leading edges of cells, as indicated in the broken line in the merged images. c, d Graphs show quantification of F-actin fluorescence intensity at lamellipodial regions (c) and at cell cortex, cytosol and overall (i.e., whole cell) (d) in MCF7-control, MCF7-K80, LTED-control and LTED-sh^a cells (n = 19, MCF7; n = 20, MCF7-K80; n = 14, LTED; n = 16, LTED-sh^a individual cells). e Graph shows quantification of percentage of cells with clear lamellipodia and membrane ruffles in MCF7-control, MCF7-K80, LTED-control, LTED-sh^a and LTED-sh^b cells (n = 8, MCF7; n = 12, MCF7-K80; n = 12, LTED; n = 7, LTED-sh^a; n = 6, LTED-sh^b fields of view). f Representative confocal microscopy images showing F-actin (magenta), pY118-Paxillin (green) and DAPI (blue) staining of MCF7-control, MCF7-K80, LTED-control, LTED-sh^a and LTED-sh^b cell. Scale bars, 25 μm. Graphs show quantification of individual leading-edge focal adhesion size (left) and number of adhesions per cell (right). Focal adhesion size (n = 269, MCF7; n = 251, MCF7-K80; n = 257, LTED; n = 331, LTED-sh^a; n = 276, LTED-sh^b). Focal adhesion number (n = 20, MCF7; n = 20, MCF7-K80; n = 20, LTED; n = 20, LTED-sh^a; n = 20, LTED-sh^b, individual cells). Statistical analyses were performed using one-way ANOVA with Tukey’s post-test. Floating bars and lines represent mean, inter-quantile distribution and SD. Asterisks represent significance at *p < 0.05, **p < 0.01 and ***p < 0.001 levels