Nature | Vol 582 | 25 June 2020 | 585retrograde actin flow to the surrounding and can therefore operate
alternately or even simultaneously. This endows amoeboid cells with
enormous adaptability when migrating in heterogeneous environments.
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© The Author(s), under exclusive licence to Springer Nature Limited 2020ControlTalin KO6–12μm (^24) smoothμm to
Migration
No migration
0
20
40
60
80
100
Talin KO cells (%)
abcd
Cell velocity(μ
m min
–1)
Control Talin KO
6 μm 12 μm 24 μmSmooth 6 μm 12 μm 24 μmSmoot
(^0) h
10
20
30
0
10
20
30
Cell velocity (a.u.)
0 1
Cell
Actin ow
0
5
10
15
20
25
Talin KO
6 μm 12 μm 24 μm
Velocity(μm min
–1)
Channel
zone
**
NS
- Smooth
ef g
31:20
00:00
(min:s)Time00:00 07:30 15:00 22:3030:00
Lifeact-GFP TIRFBright eld
6 μm 12 μm 24 μm Smooth
Channel zone smooth
Lifeact-GFP
Bright eld
Channel zone 12 μm
Lifeact-GFP
Bright eld
Channel zone 24 μm
Lifeact-GFP
Bright eld
Channel zone 6 μm
Lifeact-GFP
Bright eld
Channel
zone
Serrated Smooth
5 μm
6 μm 12 μm 24 μm **
NS
NS *
Fig. 4 | Force transmission of T cells in varying geometries. a, The channel
design that is used in b–g. b, The proportion of talin-KO cells migrating in 6-μm
and 12-μm zones (n = 41) and in the 24-μm zone and the smooth zone (n = 2 2).
**P < 0.0001, Fisher’s exact test. c, Snapshots of control cells (top) and
talin-KO cells in channels with a varying period. Lifeact-GFP is in red, and
Hoechst is in cyan. Scale bars, 20 μm. Bottom, tracks are colour-coded for cell
velocity. Data are representative of four and eight independent experiments,
respectively. d, The velocity of control (n = 88) and talin-KO (n = 79) cells in
zones with different periodicity. Data are representative of four and eight
independent experiments, respectively. Control: P = 0.0234; KO: P = 0.0188,
P = 0.0013 (6 μm versus 24 μm) and P = 0.001 (24 μm versus smooth),
*P < 0.0001, otherwise NS, Kruskal–Wallis test followed by post hoc Dunn’s
test. e, TIRF microscopy of a Lifeact-GFP-expressing talin-KO cell moving from
the 6-μm zone to the smooth zone. From top to bottom: scheme of the channel,
colour-coded snapshots every 7.5 min and a kymograph of Lifeact-GFP
(TIRF is shown in cyan and bright field is shown in grey). Scale bars, 20 μm.
Representative of three independent experiments. f, Bright-field imaging
of channel sectors and the corresponding kymograph of the Lifeact-GFP-
expressing talin-KO cell shown in e during 50 s. The red dashed lines indicate
cell displacement, and the cyan dashed lines indicate actin retrograde f low.
Scale bar, 20 μm. g, Actin retrograde f low and cell velocities of talin-KO cells
in different channel zones. Data are representative of four independent
experiments. n = 7. P = 0.0423 and **P = 0.0036, Kruskal–Wallis test followed
by post hoc Dunn’s test. Boxes in d and g extend from the 25th to the 75th
percentile, with the middle line showing the median and the whiskers
indicating the minimum to maximum values.