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. 2006 Mar 16;25(7):1456–1468. doi: 10.1038/sj.emboj.7601039

Figure 3.

Figure 3

Formation of FX-supported protrusive edges in IMP-depleted cells. (A) HeLa cells were transfected with Scr, IMP(1,3)A or IMP(1,3)B siRNAs, respectively, and stained with anti-phosphotyrosine antibody to reveal FXs and FA (left panel). Blowups (right panel) illustrate the characteristic convex FX-supported edge in an Scr-treated cell and a typical concave FX-depleted edge in an IMP siRNA-treated cell. (B) HeLa cells were transfected with Scr, IMP(1,3)A or IMP(1,3)B siRNAs, respectively, and incubated with 10 μM ROCK inhibitor (Y-27632) for 20 h and stained for F-actin (top row) or for phosphotyrosine (middle row). An overlay of these is shown in the bottom row. Scale bar, 50 μm. (C) Scatter plot of the 2D area of ROCK inhibitor-treated cells following IMP depletion. (D) Scatter plot of the roundness score from ROCK inhibitor-treated cells following IMP depletion. Cells were scored according to the formula (length of circumference)2/(4π*cell area), which is 1 for a circle (blue line). The more a cell differs from a circular shape, the higher the score. (E) Scatter plot showing the percentage of protrusive edge in IMP-depleted cells compared to Scr siRNA-treated cells. Each point in panels (C)–(E) represents a cell and the medians are indicated (red line). Asterisks depict statistically significant differences between the indicated groups, by a Mann–Whitney test (***P<0.001). The number of cells measured or scored in the groups were—Scr: n=107, IMP(1,3)A: n=122 and IMP(1,3)B: n=125.