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Figure 4


Fig. 4. Depletion of p150Glued, EB1 or CLIP-170 disrupts polarization of cells in response to scratch wounding. A549 cells transfected with siRNAs that target lamin A/C, p150Glued, EB1 or CLIP-170 were grown for 72 hours and subsequently scratched to wound the monolayer. After a further 2-hour incubation, the position of the Golgi and centrosome was determined. (A) Example wounded monolayer (lamin A/C siRNA) in which cells have reoriented towards the wound edge (white line). Also shown are dots showing the calculated cell centroid and the mask used to score reorientation. Area shown, 86x69 µm. (B) Percentage of cells showing orientation towards the wound edge was scored for each siRNA as well as for a mock-transfected control and also a scratch control (in which cells were scratched and immediately fixed). Notice that a `random' orientation, as found in the scratch control, would mean 33% of cells polarized towards the edge. Data from three experiments, >100 cells for each transfection are shown; error bars show the standard error of the mean (s.e.m.); *, significance determined using ANOVA (P<0.05).





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