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Fig. 6. Effect of silencing Pfn1 expression on single cell migration: (A) A typical time-lapse imaging experiment shows directed migration of control cells (denoted by `C') involving directed protrusion and significant net cell translocation. By contrast, Pfn1-deficient cells (denoted by `P') produce small, randomly directed protrusion with much less net cell translocation (the direction of protrusion is indicated by the arrow). Bar, 30 µm. (B) Trajectories of individual cells from the frame-by-frame analyses of the centroid of cell nuclei show a significantly (P<0.001) larger net distance traveled by the control cells (43.6±26.0 µm) compared with the Pfn1-deficient cells (11.3±9.6 µm) during the 90-minute observation period (migration data of 27 control and 21 Pfn1-deficient cells from a total of three independent experiments were pooled for the analysis). (C) Representative plots of change in protrusion direction between successive image frames (
) from a typical experiment are shown side-by-side to display much larger oscillation of in Pfn1-deficient cells (these plots were generated based on motility data of five control and six Pfn1-deficient cells in one experiment). (D) The bar graph compares the standard deviation values of for the entire 90-minute observation period (these data are based on the average values calculated for 14 control and 19 Pfn1-deficient cells - those cells that either barely protruded or came in contact with a neighboring cell at any point during the course of the experiment were excluded from the analysis). The asterisk indicates P<0.0001.
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