DOCK11 is needed for human leukocytes migration under confinement. Analysis of human HD and DOCK11-deficient T-cell migration in fibronectin-coated microchannels. Boxes include the 80% of the points, and bars represent the higher and lower 10% of points. (A) Representative montage of the change of position over time of human T-cell blast migrating inside microchannels of 4 × 5 μm with a timelapse of 2 minutes. (B) Representative kymograph of T-cell blast from HD (top panel) and DOCK11 patients (A-E, lower panels as indicated). (C) Mean instantaneous speed of HD and DOCK11 T-cell blasts migrating in 4 × 5 μm microchannels. Results from n = 2 independent experiments with each condition. Unpaired nonparametric Mann-Whitney test was used to evaluate statistical significance between the mean speeds of cells from controls and patients (∗P < .05; ∗∗P < .01; ∗∗∗P < .001). (D) Density maps representing the enrichment of actin in HD and DOCK11-deficient T cells migrating in 8 × 5 μm microchannels. Cells were allowed to migrate, fixed with 4% paraformaldehyde (PFA) to maintain the polarized morphology, and stained with phalloidin to visualize F-actin. The density maps were generated by averaging the signal from HD (n = 85), patient A (n = 32), patient D (n = 46), and patient E (n = 63) cells. Quantification of the signal intensity on density maps presented as front/rear ratio per cell. One-way ANOVA was used to evaluate statistical significance. (E) Mean instantaneous speed of HD and DOCK11 T-cell blasts (patient G) migrating in 4 × 5 μm microchannels, with or without RAC inhibitor treatment. Results from n = 2 independent experiment with each condition. One-way ANOVA test was used to evaluate statistical significance. Unpaired nonparametric Mann-Whitney test was used to evaluate statistical significance between the mean speeds of cells from controls and treated controls, patients and patients who were treated (∗P < .05).