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Figure 2


Fig. 2. Reduced lamellipodial dynamics in IQGAP1-depleted cells. Individual cells in sparse cultures that were serum-starved for 18 hours were imaged by phase contrast, time-lapse microscopy for 5 minutes before FGF2 was added to a final concentration of 25 ng/ml, and for 10 minutes thereafter. (A) Kymographic images (right panels) obtained from the indicated regions of interest (roi) at the margins of individual cells (left panels) treated with scrRNA or siRNA. Note how dynamic and motile the cell margin was in the control, scrRNA-treated cells compared to the cell depleted of IQGAP1 with siRNA. (B) Comparative responses of scrRNA-treated control (–) and IQGAP1 siRNA-treated (+) cells to FGF2 stimulation. The three parameters of lamellipodial dynamics that were measured before and after FGF2 stimulation were frequency, velocity and persistence of protrusion. The raw data were obtained from 180 regions of interest (roi) in 19 scrRNA-treated control cells, and from 180 roi in 21 siRNA-treated cells. Error bars indicate s.e.m. Differences between groups were analyzed using a one-way ANOVA test. Statistically significant differences at {alpha}=0.001 are indicated by * for control versus siRNA-treated cells after FGF2 exposure (see supplementary material Table S1 for detailed statistics, including post-hoc comparisons). The net conclusion is that control cells, but not IQGAP1-deficient cells, respond to FGF2 by making more dynamic lamellipodia.





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