Impaired cutaneous DC migration in mDia1^−/− mice. (A) The number of cutaneous DCs migrating from the skin to draining LNs. The numbers of FITC⁺ MHC class II⁺ total DCs (A) and FITC⁺ MHC class II⁺ Langerin⁺ DC and FITC⁺ MHC class II⁺ Langerin⁻ DC subsets (B) in draining LNs of B6 (open bars) and mDia1^−/− mice (shaded bars) were analyzed 72 hours after application of FITC to the footpad. Bars represent the mean ± SD from at least 3 mice per group. (C) Mobility of LCs to CCL21. Epidermal cell suspensions were applied to the upper chamber of a transwell with 3 μm (left panel) and 8 μm (right panel) pore size without coating for 3 hours. Percentages of MHC class II⁺ LCs that migrated from the upper chambers to the lower are shown. (D-E) Invasive movement of LCs to CCL21. Epidermal cell suspensions were applied to the upper chamber of a transwell coated with Matrigel for 3 hours (D) and 24 hours (E). The numbers of MHC class II⁺ cells in the lower chamber (left panels) or in the Matrigel (right panels) were counted with flow cytometry. Data are mean ± SD of 3 independent experiments. *P < .05 vs corresponding B6 mice. (F) TAXIScan assay. BMDCs chemotaxing under the CCL21 gradient were analyzed with TAXIScan. BMDCs from B6 and mDia1^−/− mice were compared in terms of the velocity. Data are mean ± SD of 4 independent experiments. *P < .05 vs corresponding B6 mice.