Fig. 3: LGR5- clusters are rounder, migrate faster and exert less traction.

a Scheme of the preparation of PDOs for cluster analysis on 3kPa 2D substrates. b Time lapse of LGR5^low and LGR5^high clusters (first and second rows, respectively). Bottom rows show the time lapse of the traction stress field of the clusters shown in the top rows. Representative images of n ≥ 24 clusters/ subgroup from four independent experiments. Scale bars, 50 μm. Tdtomato fluorescence intensity (c), cluster roundness (d), migration speed (e), and mean traction (f). Data are represented as the mean ± s.d. of n = 128 clusters from four independent experiments. Statistical significance was determined using Shapiro-Wilk normality test, followed by a Kruskal−Wallis multiple-comparison test. g LGR5^low, LGR5^med and LGR5^high clusters labeled with Sir-Actin and their corresponding XZ and YZ lateral planes. Scale bars, 50 μm. Representative images of two independent experiments. n = 49 clusters. h Mean contact angle θ between the cell cluster and the 3 kPa substrate in LGR5^low, LGR5^med and LGR5^high clusters. Data are represented as the mean ± s.d. of n ≥ 6 clusters/ subgroup from two independent experiments. Statistical significance was determined using one-way analysis of variance, followed by a Šidák multiple-comparison test. i 3D rendering of the same clusters in (g). Source data are provided as a Source Data file.