Figure 1

Overexpression of fascin leads to reorganization of actin cytoskeleton. (A) Western blot and confocal analysis of stable overexpression of Fascin-GFP (AW-Fascin-1 and AW-Fascin-2) and empty vector control pEGFP (AW-GFP-Cont) clones derived from AW13516 cells with antibodies to GFP and fascin. β-actin was used as loading control. Scale bars: 10 μm. (B) Representative confocal images of stably expressed GFP tagged fascin and GFP alone in AW-Fascin-1, AW-Fascin-2 and AW-GFP-Cont clones respectively. Cells were counter stained with DAPI. Scale bars: 10 μm. Arrow heads and arrows indicate filopodia and lamellipodia respectively. (C) Representative confocal images of F-actin stained with phalloidin-TRITC and the co-localization of F-actin with Fascin-GFP in stable AW-Fascin-1, AW-Fascin-2 and AW-GFP-Cont clones. Cells were counter stained with DAPI. Scale bars: 10 μm. Arrows indicate colocalization of Fascin-GFP with F-actin at filopodia like structures. (D) Histogram showing number of filopodia formed by fascin overexpressed (AW-Fascin-1 and AW-Fascin-2) and vector control (AW-GFP-Cont) cells. Mean and standard deviation of 3 independent experiments is plotted (p < 0.0001). (E) Histogram showing number of lamellipodia formed by fascin overexpressed (AW-Fascin-1 and AW-Fascin-2) and vector control (AW-GFP-Cont) cells. Mean and standard deviation of 3 independent experiments is plotted (p < 0.0001)