Extended Data Fig. 6: Topography-based migration of primary T cells.
a, Scheme (top and side view) of control and Lifeact-GFP cells confined under agarose on a smooth surface coated with Fc-ICAM1 or passivated with PLL-PEG. WT, wild type. b, Scheme (top and side view) of control and Lifeact-GFP cells confined under agarose on a coverslip with linear ridges passivated with PLL-PEG. c, Minimum-intensity projection showing the trace of control cells migrating for 15 min under agarose on Fc-ICAM1-coated smooth surfaces (left), PLL-PEG-coated smooth surfaces (middle) and PLL-PEG-coated ridge-baring surfaces (right). Representative of six independent experiments. Scale bars, 50 μm. d, Randomly selected tracks of mixed control and Lifeact-GFP cells migrating under agarose on Fc-ICAM1-coated smooth surfaces (left, n = 183), PLL-PEG-coated smooth surfaces (middle, n = 183) and PLL-PEG-coated ridge-baring surfaces (right, n = 183). The orientation of ridges is indicated in the top right corner. e, Rose diagram (with circular lines from the centre showing 6%, 12% and 18% of total cells) of cells migrating on Fc-ICAM1-coated smooth surfaces (left, n = 419), PLL-PEG-coated smooth surfaces (middle, n = 650) and PLL-PEG-coated ridge-baring surfaces (right, n = 687). The orientation of ridges is indicated in the top right corner. n = 6 independent experiments. f, g, Velocities of cells migrating under agarose on Fc-ICAM1-coated smooth surfaces (n = 419), PLL-PEG-coated smooth surfaces (n = 650) and PLL-PEG-coated ridge-baring surfaces (n = 687). n = 6 independent experiments. Cell velocities (cell displacement divided by track duration) (f) and mean instantaneous velocities (g) are shown. ****P < 0.0001 and ns P = 0.0757, both one-way ANOVA with Kruskal–Wallis test followed by post hoc Dunn’s test.
