Extended Data Fig. 4: Effect of topographical features on T cell migration.
a, Related to Fig. 4d. Scheme of channel designs with decreasing complexity. In brief, serrations with a period of 6, 12 and 24 μm were arranged in consecutive zones of a 5-μm wide channel ending with a smooth zone. The left panel shows the different geometries. The right and middle panels show the velocity of cells migrating in zones with a different period. The total number of cells in each design is shown. Representative of four (control cells) and eight (talin-KO cells) independent experiments. Control: *P = 0.0415. KO arrow ‘right’: ***P = 0.0008 and **P = 0.0013; KO arrow ‘left’: ***P = 0.0003 and **P = 0.0018; KO bulb: *P = 0.0344; KO half-bulb: **P = 0.0016 (6 μm versus smooth) and **P = 0.0013 (12 μm versus smooth), otherwise not significant, one-way ANOVA with Kruskal–Wallis test followed by post hoc Dunn’s test. Boxes extend from the 25th to the 75th percentiles, with the middle line showing the median and the whiskers representing the minimum to maximum values. b, Top, scheme of cell and actin retrograde flow length. Bottom, control cells expressing the Lifeact-GFP reporter with 10 mM EDTA (grey) or talin-KO cells (red), under 5-μm confinement (left, n = 12) or in a 5 × 5-μm channel (right, n = 9 for control cells and n = 8 for talin-KO cells), were observed by TIRF microscopy, and the mean cell length (ns P = 0.0555, two-sided Mann–Whitney U-test) and the actin retrograde flow length (ns P = 0.7247, two-sided Mann–Whitney U-test) were measured using kymograph analysis. Representative of two independent experiments. Data are mean ± s.d.
