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. 2021 Mar 25;220(6):e202007182. doi: 10.1083/jcb.202007182

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© 2021 Robertson et al.

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Figure 6. — ERM proteins are required for S1P-induced chemokinetic responses. (a–h) Cells were cultured overnight in media with 10% CS-FBS, added to VCAM-coated chambers, and monitored by DIC microscopy following stimulation with S1P (100 nM), CCL19 (100 ng/ml), or CXCL12 (100 ng/ml). (a and b) Percentage of cells that displace at least 10 μm during the response. Each dot corresponds to a field of cells pooled from three independent experiments shown. Means ± SD are shown and compared individually using t tests (two-sided). (c and d) Displacement of WT and DKO migratory cells following S1P or chemokine stimulation. Dashed lines indicate the 10-μm threshold. (e and f) Directionality of migratory WT and DKO cells. Directionality was calculated by dividing displacement by total track length. (c–f) Pooled data from three independent experiments are displayed as individual cells (dots) and means (horizontal bars) and compared using Mann-Whitney (nonparametric) t tests. (g and h) Tracks of individual cells with the same origin are shown, representative of three individual experiments. n.s., P > 0.05; *, P < 0.05; **, P < 0.01; ****, P < 0.0001.