Figure 2

MEC1 are suitable for analysis of directional chemotaxis. (A) Dunn Chamber: the outer pool is filled with medium with/without chemokine, the inner pool is filled with medium only. Cells are scanned on the bridge between outer and inner pool. (B) Four graphs describe chemotaxis of MEC1 cells in a Dunn Chamber. Control (CTRL) - no chemokine gradient between two pools (n = 3). CCL19 gradient between inner and outer pool (n = 5). CK1 inhibition shows a condition with chemokine gradient between the pools in combination with CK1 inhibitor PF670642 (n = 3). Migration properties of tracked MEC1 cells are defined by Track length (exact distance of cell path), Track Displacement (distance of shortest path between each cell’s starting and the end point), Maximum Speed (maximum reached speed of each cell during the whole measurement) and Straightness (absolutely straight path = 1, minimum = 0). Statistical analysis was performed in GraphPad, using the Kruskal-Wallis test. (***, P < 0.001). (C) The representative example of trajectories of MEC1 cells in Dunn chamber-generated CCL19 gradient. Cells were treated with control solution (up) or CK1 inhibitor (bottom). The source of chemokine is indicated by a star. White arrows indicated the direction of migration by connecting the start and end point. (D) The statistical analysis of the directionality of migration was visualized using Oriana software. Individual blue dots (corresponding to individual cells) are plotted on the circle based on the deviation of their migration direction from the CCL19 source, which is set as 0° (up). Data were analyzed by the Rayleigh Test for randomness of circular data. Arrow points towards the predominant direction of cell migration; arrow length indicates statistical significance where only arrows crossing the inner circle (representing p = 0.05) are statistically significant. Please note that CCL19 adds a strong directional component to the migration, which is lost following CK1 inhibitor treatment. ***, P < 0.001.