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

Figure 4.

Figure 4

Inhibition of MDA-MB-231 OXPHOS lowers cellular energetics and cytoskeletal activity in 3D collagen gels. (A) A color-coded time-lapse of MDA-MB-231 protrusive activity in collagen matrices is shown (dashed yellow squares show zoomed regions). Quantification of (B) stepwise cell velocity (n = 33–61 cells) and (C) stepwise cell protrusive activity (n = 30 cells) in matrices of increasing collagen density is shown. (D) Normalized PercevalHR ratio of MDA-MB-231 cells in collagen matrices is shown (yellow lines show cell outlines). Quantification of (E) normalized PercevalHR ratio (n = 33–61 cells) and (F) TMRM fluorescent signal (n = 48–93 cells) in matrices of increasing collagen density is shown. Quantification of (G) normalized PercevalHR ratio (n = 32–61 cells), (H) TMRM fluorescent signal (n = 48–93 cells), (I) stepwise velocity (n = 32–61 cells), and (J) protrusive activity (n = 30 cells) after AMA treatment in matrices of increasing collagen density is shown. (K) Percentage change in cellular energetics, migration speed, and protrusive activity averaged for each collagen density after AMA treatment in comparison to control conditions 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; dashed lines show linear regression; two-tailed t-test was used when comparing between ctrl and AMA, and multiple comparisons with ANOVA was used when comparing across collagen densities; n.s., nonsignificant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. Scale bars, 50 μm. To see this figure in color, go online.