Figure 1

Analysis of B lymphocyte (MEC1) cell migration by live cell imaging. (A) A photomicrograph showing strongly polarized migrating MEC1 cells with the clearly defined leading and trailing edge. Arrows indicate the direction of migration. Size bar = 10 μm. (B) Snapshots of migrating, cell tracker-stained, MEC1 cells from time-lapse microscopy at approx. time: 0, 5, 10 and 15 min. Individual moving cells are indicated by the same color-coded arrow in each snapshot. Size bar = 10 μm. (C) MEC1 cells were seeded on glass-bottom plates coated with human plasma fibronectin. Control condition was captured first, for 30 min. Subsequently, CCL19 chemokine was added to the cells and the same position was scanned for additional 30 min. Last, cells were inhibited with PF670462 and followed another 30 min. (D) Four-compartment glass bottom plates were used for parallel tracking the cells (control [1], CCL19-treated [2] and CCL19/CK1 inhibitor-treated [3]). (E) Ibidi chamber for self-insertion was inserted in a glass-bottom plate, coated with Fibronectin and cell-tracker stained MEC1 cells were seeded. Ibidi chambers allow parallel scanning in two conditions (control [1a] and CCL19-stimulated [2a] first, captured for 30 min). After that, cells were stimulated with the chemokine [1b] and one of the pools was inhibited with the CK1 inh I [2b] and scanned for additional 30 min. From the first period, MEC1 migration in control and CCL19-stimulated wells [1a, 2a] is compared to each other and after that, CCL19 only and CCL19 + CK1 inh [1b, 2b] are compared to each other. At least 50 cells was tracked in each condition. ***, P < 0.001 (Kruskal-Wallis test).