Actin cortexCell lengthTIRF Lifeact-GFPTIRF Actin
retrograde flowa6-m
12-m
24-m
Smooth01020306-m
12-m
24-m
Smooth01020306-m
12-m
24-m
Smooth01020306-m
12-m
24-m
Smooth01020306-m
12-m
24-m
Smooth01020306-m
12-m
24-m
Smooth01020306-m
12-m
24-m
Smooth01020306-m
12-m
24-m
Smooth0102030Cell velocity (μm/min)Cell velocity (μm/min)Cell velocity (μm/min)Cell velocity (μm/min)Control Talin KO b**************** 2.5D14μm27μmControl + EDTA
Talin KOActin retrogradeflow length (μm)Smooth
channel01020300204060800204060800102030Cell length (μm)Control + EDTAns
17μm19μm39μm49μmns5μm 5μm5μmn=28n=13n=15n=31n=28n=15n=22n=15Extended 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 f low 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 f low 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.