Fig. 2: Different modes of migration and mitochondrial metabolism.

a, b Oxygen consumption rate (OCR) from cells embedded in a 3D collagen I matrix (n = 3). Comparison between elongated-mesenchymal and rounded-amoeboid cells in a panel of cell lines and between the melanoma pairs. c Maximal intensity projection (left) and 3D representation (right) of ATP:ADP ratio in HT1080 and A375M2 cells expressing Perceval HR biosensor, embedded in a 3D collagen I matrix. Scale bar = 10 μm. d Quantification of ATP:ADP ratio from (c) (18 cells/condition pooled from n = 3). e AMP levels quantified by ¹H-NMR for the indicated cell lines (n = 3). f Intracellular ATP levels in WM983A and A375P cells grown under adherent or floating conditions for 24 h (WM983A, n = 4; A375P, n = 5). g XF ATP rate index indicative of the ratio of mitochondrial and glycolytic ATP Production Rate in A375P under adherent or floating conditions for 24 h (n = 4). h Quantification of Tetramethylrhodamine Ethyl Ester Perchlorate (TMRE) fluorescence intensity normalized by MitoTracker Deep Red signal by FACS in cells adhered on plastic or floating cells for 24 h. Data shown as fold versus adherent cells (n = 3). Graphs (a, b, f, g, h) show mean ± SEM. Box plots (d, e) show median (centre line), interquartile range (box) and min-max values (whiskers). p values were calculated using two-tailed tests (a, b, d–g). p value by unpaired t-test comparing elongated-mesenchymal versus rounded-amoeboid cells (a), unpaired t-test (b), paired t-test (e–g), Mann-Whitney test (d) and two-way ANOVA with Sidak’s correction comparing floating vs adherent cells (h). All n are indicative of independent experiments unless otherwise stated. Source data are provided as a Source Data file. See also Supplementary Fig. 2.