Fig. 1: Cytoskeleton, adhesion and traction stress in 3D migration. | Nature Communications

Fig. 1: Cytoskeleton, adhesion and traction stress in 3D migration.

From: AMPK is a mechano-metabolic sensor linking cell adhesion and mitochondrial dynamics to Myosin-dependent cell migration

Fig. 1

Cells seeded on a 3D collagen I matrix. a pMLC2 and F-actin confocal images in HT1080 and A375M2 cells (n = 3). Scale bar = 50 μm. b, c Quantification of cell morphology (211, 291, 313, 358, 339 and 311 cells pooled from n = 6) (b) and pMLC2 immunofluorescence signal normalized by cell area (45, 72, 65, 79, 85 and 83 pooled from n = 3) (c). d Representative fluorescence intensity line scans (dashed white lines in image) showing distribution of F-actin (red), pMLC2 (green) and nucleus (blue) along elongated-mesenchymal (HT1080 and A375P) and rounded-amoeboid (A375M2) cells (n = 10 cells/cell line). e Quantification of points of attachment between cells and matrix (78, 99, 61 and 112 cells pooled from n = 3). f Percentage of adhered cells 1 h after seeding on a matrix of collagen I (n = 4). g Scheme showing that cell morphology, Myosin II activity and adhesion to the matrix define the mode of migration and the stress exerted into the matrix (Created with BioRender.com). h, i Cells treated with vehicle (DMSO) or blebbistatin (25 μM). (Left) Representative displacement vector maps from maximum intensity projections of cells obtained from the tracked displacements of collagen I fibres between 0 to 2 min. Red and green boundaries indicate the outline of the cell in the previous and current frames, respectively. Scale bar = 20 μm. (Right) Representative traction stress magnitude maps corresponding to displacement maps. Colour bar indicates traction stress magnitude (Pascal, Pa). Scale bar = 20 μm. Dot plot (e) shows median with interquartile range (each dot represents a single cell). Violin plot (b) shows median with interquartile range. Box plots (c) show median (centre line), interquartile range (box) and min-max values (whiskers). Graph (f) shows mean ± SEM. p values by Kruskal–Wallis test with Dunn’s multiple comparisons test (b, c, e), one-way ANOVA with Dunnett’s correction versus A375M2 (f). All n are indicative of independent experiments unless otherwise stated. Source data are provided as a Source Data file. See also Supplementary Fig. 1.

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