Fig 1: Analysis of motility and directionality of cell islands composed of tight junction protein knockdown cell lines and comparison with AGS and EC96. (a) Bright-field images of CLDN7 and ZO-1 KD cell islands at 0 h and 9 h after release (scale bar = 200 µm). (b) Relative area increase of cell islands composed of the AGS (blue circle), EC96 (red square), CLDN7 KD (cyan triangle), and ZO-1 KD (green diamond) cells over time. The error bars represent the standard deviations calculated from separate assays on each group (n = 5). (c) The trajectory of the cells within the CLDN7 KD and ZO-1 KD cell islands. The stronger red color of the arrows indicates more time passed. (d) Average path length along the same distance from center to edge of the AGS (blue circle), EC96 (red square), CLDN7 KD (cyan triangle), and ZO-1 KD (green diamond) cell islands. The error bars represent the standard deviations calculated from separate assays on each group (n = 5). (e) Color-code map of the directional persistence of the cells within the CLDN7 KD and ZO-1 KD cell islands. The brighter dots indicate higher directional persistence. (f) Average directional persistence along the same distance from center to edge of the AGS (blue circle), EC96 (red square), CLDN7 KD (cyan triangle), and ZO-1 KD (green diamond) cell islands. The error bars represent the standard deviations calculated from separate assays on each group (n = 5). (g) Color-code map of the directional persistence of the low 25% (gray edge circles) and high 25% (black edge circles) of motile cells within the CLDN7 KD and ZO-1 KD cell islands. Gray circles represent mid-quartile (25–75%) motile cells within each group population. (h) Average directional persistence of low and high motility cells within the AGS (blue), EC96 (red), CLDN7 KD (cyan), and ZO-1 KD (green) cell islands. The error bars represent the standard errors calculated from separate assays on each group (n.s. not significant, * p < 0.05 and *** p < 0.001; n = 5).