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. 2021 Oct 14;11:20434. doi: 10.1038/s41598-021-99902-9

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© The Author(s) 2021, corrected publication 2021

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Cancer cell migration in 3D ECM is coupled with morphological phenotype and phenotype transitions. (A) Means (m, upper panel) and variances ( $σ^{2}$ , lower panel) of the step size distributions by fitting the experimental measurements with log-normal distribution. The steps are categorized based on morphological phenotype (coarse-grained as mesenchymal, amoeboidal and intermediate state) dynamics. ME: abbreviation for mesenchymal, which includes FP and LA states. AM: abbreviation for amoeboidal, which includes BB and AE states. If a cell makes a step by starting from AM state to ME state, the step is categorized as an AM-ME step. The starting and ending frames of steps are separated by 15 minutes. In (A) errorbars show the 95% confidence interval of fitted parameters. (B) Real-space (2D projection) mean square displacements (MSD) of cells. AM: the MSD of cells dwelling in the AM state. ME: the MSD of cells dwelling in the ME state. average: the weighted average of mean square displacements of AM, ME, and intermediate dwelling cells. The average is based on the occurrence fraction of AM, ME and intermediate state of cells. Full trajectory: the MSD obtained from entire cell trajectories regardless of the morphological phenotypes. Shaded areas in C show SEM. In (A-B) a total of 1974 hours of single cell trajectories are analyzed. The ECM are prepared at room-temperature with a concentration [col] = 1.5 mg/mL. This figure is prepared with Matlab R2020a (www.mathworks.com) and ImageJ (https://imagej.net).