Fig. 2. Cells assayed for migratory behaviour in Dunn chambers. (A) Growth medium
(+ 10% FCS) without chemoattractant gradient. (B-D) starvation medium (+ 0.1%
FCS). (B) No chemoattractant gradient. (C) Concentration gradient of EGF (15
ng/ml in outer well). (D) Concentration gradient of mut C uPA (10 nM in outer
well). (A-D) Cell trajectories over 5 hours (10 median cell trajectories). The
start point for each cell is coordinate (0,0); The end point is marked by a
closed circle; the chemoattractant source is at the top of plot; axis
graduations, distance (µm). Circular histograms show the proportion of
cells with migratory direction lying within each 20° interval
(chemoattractant source at top of histogram). Cells that migrated less than 20
µm were excluded from analysis. The red arrow represents the mean direction
of migration; the green segment represents the 95% confidence interval
determined by the Rayleigh test. Displacement shows the percentage of cells
that migrate further than 20 µm. Concentration-dependent response for (E)
EGF, (F) wt uPA (open circles) and mut C uPA (closed circles). (G) Effect of
ß1-integrin neutralising antibody and mouse IgG antibody on
chemotaxis stimulated by 10 nM mut C. More than 40 cells pooled from three to
six experiments included in analyses.