Figure 6
Mena11a expression decreases G-actin incorporation to F-actin barbed ends at the leading edge.
All experiments done on MTLn3 cells stably expressing GFP, Mena and Mena11a. (A,D,G): Barbed end incorporation after stimulation with (A) 0.5 nM EGF for 60 seconds, (D) 5 nM EGF for 60 seconds, and (G) 5 nM EGF for 180 seconds. Barbed ends and F-actin visualized with rhodamine-G-actin and phalloidin labeling, respectively. Scale bar, 10 μm. Insets at (A) 27X, (D) 31X, and (G) 25X magnification show barbed end distribution at the leading edge. (B,E,H): Quantification of relative number of barbed ends at leading edge, after stimulation with (B) 0.5 nM EGF for 60 seconds, (E) 5 nM EGF for 60 seconds, and (H) 5 nM EGF for 180 seconds. Error bars: SEM. Results represent triplicates, >30 cells analyzed for (B,E) >50 cells for (H). One-way ANOVA **p < 0.01, ***p < 0.005, n.s not significant. (C,F,I): Normalized pixel intensities of relative number of barbed ends, plotted as a function of distance from the cell edge (mean ± SEM), after stimulation with (C) 0.5 nM EGF for 60 seconds, (F) 5 nM EGF for 60 seconds, and (I) 5 nM EGF for 180 seconds.
