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Fig. 5. N-WASP is required for FGF2-stimulated migration of MDBK cells. (A) MDBK cells were transfected with N-WASP-specific siRNA or scrambled RNA (scrRNA) using a Nucleofector. To allow direct visual comparison of N-WASP levels in the two samples by western blotting, a concentration series of each cell extract was analyzed at the indicated relative dilutions. Note that siRNA reduced the N-WASP level to 
1/3 of normal, but had no effect on cellular IQGAP1 content. (B) Confluent monolayers were then serum-starved for 8 hours, wounded with a micropipette tip, and 2 hours later were stimulated with 25 ng/ml FGF2. Note that movement of N-WASP-depleted cells into the wound after 9 or 24 hours of FGF2 exposure was severely impaired. (C) Percentage of wound closure was quantified by measuring the average width of eight randomly chosen regions of interest of each wound at 0, 9, and 24 hours after FGF2 addition. Data are expressed relative to the average wound widths at 0 hours. Error bars indicate standard deviations for three experiments, and asterisks (*), indicate significant differences between scrRNA controls and corresponding siRNA-treated cultures at 
<0.001.
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