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Figure 3


Fig. 3. Cdc42-siRNA-treated cells show a loss of actin nucleation at the protruding edge in response to EGF stimulation. (A) Representative micrographs of MTLn3 cells transfected with control luciferase or Cdc42 siRNA, starved for 3 hours and stimulated with EGF for 1 or 3 minutes or not (0 min). Cells are stained with Rhodamine-phalloidin to visualize F-actin. (B) Quantification of the average F-actin fluorescence at the protruding edge (0.2-0.6 µm in from the edge of the cell) normalized to edge intensity of non-stimulated cells. Data are the mean ± s.e.m. from three different experiments (15 cells per experiment). (C) Control and Cdc42 siRNA-treated MTLn3 cells were stimulated with EGF for 1 or 3 minutes or not (0 min), permeabilized and incubated with biotin-actin, and immunostained with anti-biotin to detect the newly formed barbed ends. (D) Quantification of barbed-end formation at the protruding edge (0.2-0.6 µm in from the periphery) normalized to unstimulated cells. Data are the mean fluorescence ± s.e.m. from three different experiments (15 cells per experiment). Bars, 10 µm.





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