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. 2018 Nov 20;9:4891. doi: 10.1038/s41467-018-07290-y

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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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Fig. 5 — Arp2/3 suppression promotes a microtubule-dependent uniaxial guidance response. a MDA-MB-468 breast carcinoma cells under Arp2/3 complex inhibition (+CK666) sense soft (2.3 kPa) and stiff (50 kPa, Supplementary Movie 12) biaxial collagen guidance cues. Note that cell populations on stiff substrates split into vertical and horizontal cell subpopulations (V vertical cells, H horizontal cells). b Morpho-mechanical analysis of cells populations under Arp2/3 inhibition conditions on soft and stiff biaxial cues: free cell edge curvature radii, R. c Average values of R and R/d in cell under Arp2/3 complex inhibition, showing increased traction on stiffer substrates (see Fig. 2g for comparison). The corresponding n values are shown on b. d, e 3D reconstruction overviews and morphometric analysis of the Arp2/3-inhibited cell population response to biaxial guidance cues. Inactivation of Arp2/3 (+CK666) induces non-equidirectional cell protrusion with random cell commitment to either of the two orthogonal directions (i.e., a uniaxial response). Disruption of intact microtubules in cells with suppressed Arp2/3 complex (+ CK666+nocodazole) rescues biaxial cell spreading. f Average values of cell lengths from protrusion for each of the conditions in d, e (see color code). The corresponding n values are shown on e. Scale bars—30 µm. All corresponding n values are shown on the plots. Number of replicates (independent experiments) for all measurements N = 4. Data in a, c, f are mean ± s.d.; ns: no significant difference between groups; *p < 0.05, **p < 0.001 (c: unpaired t test; a, f: ANOVA)