Figure 1.

DCs display two types of migration modes into lymphatic vessels. (A) Spinning disc confocal images (maximum projection of z-stack, 22 planes, 0.5 µm distance) of labeled DCs (red) migrating into lymphatic vessels (BM stained for laminin, green) over time. See also Video 1. Bar, 50 µm. (B) Enlargement of inset in (A) representatively showing tracks of four different DCs. Bar, 20 µm. On the right of the panel instantaneous velocities of the DCs (tracks 1–4) are plotted over time. Arrival at the lymphatic vessel BM is marked by a red and appearance in the vessel lumen by a green arrow. (C) Average migration velocities in the presence of DMSO (control) or protease inhibitor cocktail (PIC). Control: 5 independent preparations, 56 cells tracked. PIC: 7 independent preparations, 61 cells tracked. Error bars show the mean ± SE of mean (SEM). ns, not significant. (D) DC migration across plain transwell (PET) vs. transwell carrying an in vitro assembled BM (BM) in the presence of DMSO (control) or PIC. One representative of two independent experiments. Each dot represents a single transwell used in the experiment. Error bars show mean ± SEM. ns, not significant. ***, P < 0.0001 (E) Confocal images of whole-mount ear skin 2 h after addition of exogenously labeled bmDCs. DCs were incubated before crawl-in assay with either DMSO (1) or protease inhibitor (2). Bar, 20 µm. Images are representatives of one of at least five independently performed crawl-in assays.