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. 2021 Mar 9;120(9):1705–1717. doi: 10.1016/j.bpj.2021.02.044

Figure 5.

Figure 5

Protrusive activity is positively correlated with cellular energetics and motility in high-density collagen matrices through OXPHOS. (A) Color-coded time-lapses of MDA-MB-231 protrusive activity and migration after AMA treatment in matrices of increasing collagen density are shown (dashed squares show zoomed regions; yellow, ctrl; red, AMA). (B) Normalized PercevalHR ratio of MDA-MB-231 cells after AMA treatment in matrices of increasing collagen density is shown (yellow lines show cell outlines). (C) Stepwise cell protrusion and total change in ATP/ADP signal of single MDA-MB-231 cell after AMA treatment in collagen matrices are shown (n = 20–40 cells). (D) Normalized PercevalHR ratio of MDA-MB-231 cells in matrices of increasing collagen density with confocal reflectance is shown (small yellow arrows show fiber directions). (E) Quantification of cell contractility and collagen fiber orientation within 10 μm to cell protrusions for MDA-MB-231 cells in matrices of increasing collagen density is shown. (F) Motile fraction as a function of stepwise new protrusions averaged for each collagen density is shown. (G) Quantification of motile fraction (n = 12–20 positions) after AMA treatment in matrices of increasing collagen density. (H) Percentage change in protrusive activity and motile fraction after AMA treatment for MDA-MB-231 cells in matrices of increasing collagen density is shown (AMA/Ctrl; no change = 1; complete inhibition = 0). Data shown as box-and-whisker plots denote mean (+) or medians and 25th/75th and 5th/95th percentiles; lines show LOWESS smoothing, and dashed lines show linear regression with mean ± SE; P represents extra-sum-of-squares F-test when comparing linear regression. n.s., nonsignificant; ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Scale bars, 50 μm. To see this figure in color, go online.