Figure 3: Cell polarization in migrating fibroblasts.
From: Novel Coronin7 interactions with Cdc42 and N-WASP regulate actin organization and Golgi morphology
(a) Schematic showing a cell with Golgi and MTOC positioned within a 120° sector, ( + ) polarized; (−) non-polarized. Blue arrow, direction of migration; black solid line, wound edge. (b–d) Scratch-wounded WT and KO fibroblasts stained for (b) Golgi (58K, green) and (c) centrosome (Pericentrin, cyan) 3 h post-wounding. Nuclei stained with DAPI (blue). The position of the leading zone (broken white line) shown. Scale bar = 25 μm. Line chart (d) showing percentage of cells with reoriented Golgi and MTOC (n = 100 cells each time, each condition, 2 independent experiments; *P < 0.05). (e) TRITC-phalloidin staining of wounded cell layers (red false-coloured as grey) to visualize the F-actin network. White arrowheads, orientation of the actin fibres (parallel in WT and perpendicular in KO). Scale bar = 50 μm. (f) Bar graph with percentage of cells at the wound edge with a polarized distribution of F-actin i.e., perpendicular to leading edge (n = 100 cells each, 2 independent experiments; **P < 0.01). (g) Analysis of cell migration over 20 h for WT and KO fibroblasts. Scale bar = 100 μm. Frames from time-lapse phase-contrast videos at the indicated time shown. (h) Analysis of the velocity of single cells (in μm/hour). 10 cells from both WT and KO, per position and per edge were considered (n > 200 cells, 5 independent experiments; **P < 0.01). Data shown as mean ± SD.
