Fig. 3
Cell stiffness alters decision-making and cell mechanics during confined migration. a AFM measurements of cell stiffness following pharmacological treatments and siCav1 knockdown (n = 164, 95, 167, 91, 75, 85, 92, and 97 cells for Ctrl, Rho+, CL-A, Y27, ML7, MβCD, siCtrl, and siCav1) and western blot for Caveolin-1 following treatment with 25 nM Caveolin-1 siRNA. b, c Migration decision-making (b) and passing time into each branch (c) following treatments to alter cell stiffness (n = 30, 34, 30, 31, 30, 28, 33, and 39 cells for Ctrl, Rho+, CL-A, Y27, ML7, MβCD, siCtrl, and siCav1). d–f Model predictions for cell deformation (d), track displacement (e), and the inverse relationship between cell elongation and track displacement (f) in 7 μm tracks. g Transmitted light images showing cell morphology and confocal reflectance images of microtrack structure as well as cell-induced track displacement in 7 μm microtracks (yellow lines show microtrack walls; red arrowheads show areas of matrix displacement around the cell body). h–j Experimental averages of cell elongation (h), track displacement (i), and the relationship between cell remodeling and matrix displacement (j) in 7 μm microtracks as a function of cell stiffness (n = 118, 107, 100, 135, 109, 123, 53, and 58 cells for Ctrl, Rho +, CL-A, Y27, ML7, MβCD, siCtrl, and siCav1). Data shown as median ± interquartile range (box), 5th–95th percentiles (whiskers), and mean (+) (a, c), or mean ± s.e.m. (b, h–j); dashed lines show one-phase decay (h, j) and one-phase association (i); two-tailed Mann–Whitney test (a, c) or two-tailed t-test (b); *P < 0.05, **P < 0.01, ***P < 0.001. Scale bar, 15 μm
