Loss of Gadkin impairs BMDC chemotaxis in 2D.

<p>A,B. LPS-matured WT and Gadkin^-/- BMDCs were seeded into Transwell inserts and allowed to migrate for 3 h towards 250 ng/ml CCL19 (A) resp. 250 ng/ml CCL21 (B). Migration towards the chemokine was analyzed by flow cytometry. Values were subtracted for unspecific migration towards chemokine-free medium to determine the specific migration towards chemokine and then normalized to the average value of each experiment (N = 6 independent experiments, unpaired two-tailed t-test, * = p<0.05). C-H. LPS-matured WT and Gadkin^-/- BMDCs were seeded into a 2D chemotaxis chamber. BMDCs were allowed to migrate for 4 h towards 650 ng/ml CCL19, and their migratory behavior was evaluated based on bright-field images obtained at 2 min intervals. C,D. Migration tracks of 21 WT (C) and 23 Gadkin^-/- BMDCs (D) centered on the same starting point. Center of mass depicted as blue cross (WT: x = 10.57 μm, y = 326.38 μm, KO: x = 4.26 μm, y = 213.79 μm). Velocity (E) was quantified as well as the accumulated (F) and Euclidian distance (G) covered by the BMDCs. Directionality (H) was calculated as Euclidian distance divided by accumulated distance (N = 4 independent experiments, at least 20 cells per experiment were tracked, paired two-tailed t-test for distances, paired one-tailed t-test for velocity, * = p<0.05).