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Fig. 1. Cofilin recruitment to the leading edge of the lamellipod in the initial phase after EGF stimulation is initiated faster than Arp2/3 recruitment. (A) CFP-Arp3 and YFP-cofilin distribution in the same cell before and after EGF stimulation. Arrowheads indicate accumulation of GFP-Arp3 at the leading edge. Bar, 10 µm. (B-D) Cells transfected with GFP-cofilin or GFP-Arp3 were time-lapsed after the addition of EGF, followed by measurement of the fluorescent edge intensity and cell area throughout the time-course. The area was plotted as fold area increase over time (B and C, open circles). The fluorescent edge intensity in a 0.7 µm depth from the edge of the membrane was plotted as mean fold fluorescent intensity increase over time (B: filled squares, GFP-Arp3, n=7; C: filled triangles, GFP-cofilin, n=9). Error bars indicate the standard error of the mean (s.e.m.). (D) To calculate the initial rates of intensity increase for Arp3 and cofilin, data points for the intensity plots in B and C from 20 to 60 seconds were fitted to straight lines and the slopes of these lines were taken as the initial rates of increase. To estimate errors, the slope calculations were conducted with the data from B and C plus or minus s.e.m.
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