Fig. 4

a, Illustration of cell preconditioning at the indicated viscosities. b, Confined migration speeds of preconditioned cells (0.77 or 8 cP for 6 days) resuspended at the indicated migration viscosity. Data are mean ± s.d. for n ≥ 140 cells from 3 experiments. c, The confined migration speeds of preconditioned SC or shTRPV4 cells allowed to migrate at 0.77 cP. Data are mean ± s.d. for n ≥ 146 cells from 3 experiments. d, Confocal image of 3 day post-fertilization (d.p.f.) zebrafish ISVs with measurements of vessel width (top). Bottom, experimental design of migration studies in zebrafish. e,f, Time-lapse confocal images (e) and average speeds (f) of preconditioned cells (n ≥ 77) inside ISVs from 3 experiments. The red lines indicate the median (thick) and quartiles (thin). g, The experimental design of mouse tail-vein experiments. h, The number of human vimentin-positive colonies in the lungs 48 h after injection. Data are mean ± s.e.m. for 8 mice per group from 2 experiments. i,j, Confocal images of lung sections (i) and quantification of human vimentin-positive metastatic colonies (j) 3 weeks after injection. Data are mean ± s.e.m. for a total of ≥7 mice per group from 2 experiments. k,l, The number of human vimentin-positive metastatic colonies in the lungs 48 h (k) and 3 weeks (l) after injection. Data are mean ± s.e.m. for ≥9 mice per group from 2 experiments. The squares represent experiments with PVP as the medium additive. m,n, qPCR detection of human DNA in the lungs of mice 48 h (m) or 3 weeks (n) after injection. Data are mean ± s.d. for ≥9 mice per group. Squares are from experiments with PVP. Statistical analysis was performed using Kruskal–Wallis tests followed by Dunn’s test (b and c), Mann–Whitney U-tests (f), unpaired t-tests (h and j), one-way ANOVA followed by Tukey’s test (l–n) and one-way ANOVA followed by Tukey’s test on log-transformed data (k). Scale bars, 20 µm (d), 30 µm (e) and 200 µm (i). The schematics in a, d and g were created using Servier Medical Art.

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