Figure 4

Cdc42 blockade reduces colon cancer cell migration, invasion and affects actin cytoskeleton reorganisation. A Representative images of migrated SW620 colon cancer cells from an in vitro migration assay are shown. Colon cancer cells were treated with 1, 2 or 5 μM AZA197 for 24 h and migrated cancer cells quantified subsequently by in vitro migration assays. Data were collected from five individual consecutive fields of view (40x) from three replicate Boyden chambers. *, significantly different from controls. B The invasive capacity of SW620 cells was determined in matrigel invasion assays. Invaded SW620 cells were quantified from five individual consecutive fields of view (100x) from three replicate chambers. *, significantly different from control. C Effect of AZA197 treatment on cell morphology, filopodia formation and actin reorganization. SW620 colon cancer cells were plated on fibronectin/gelatin coated cell culture chambers and incubated with 2, 5 and 10 μM AZA197 for 24 h. Paraformaldehyde fixed cells were stained with Atto-488 phalloidin (F-actin, green) to visualize the polymerized actin cytoskeleton and filopodia and subsequently counterstained with DAPI (blue) and photographed (magnification, x1000). Arrows indicate filopodia. AZA197 leads to changes in cellular morphology and suppresses filopodia formation.