Extended Data Figure 7. Wnt enhances motility in vitro.

a, Percentage of fast-moving (>2 μm/min), slow-moving (0.3-2 μm/min) and non-moving (<0.3 μm/min) Lgr5⁺ cells. The imaged Lgr5⁺ cells were isolated from Lgr5-EGFP-ires-creERT2;R26R-confetti organoids and exposed to (I) control medium (n=408 cells), (II) medium supplemented with Wnt3a (n=582 cells), (III) medium supplemented with Paneth cells (PC) (n=418 cells), or (IV) medium supplemented with PC and Wnt inhibitor (IWP2) (n=431 cells) in Matrigel from 3 independent biological replicates. b,c Speed (b) and directionality ratio (persistence) over time calculated as mean displacement/length of the trajectory. Significance was determined by a two-sided Mann-Whitney test.(c) of single Lgr5⁺ cells in control medium, medium supplemented with Wnt3a, co-culture with PC and co-culture PC with IWP2. Shown are n=150 random cell tracks of Lgr5⁺ cells from 2 independent organoid lines, 50 from each of 3 independent biological replicates. Each point represents the mean value of each track. Shown are mean ± SEM.