Figure 4

CCN5 targeted siRNA promotes motility in growth arrested VSMC. VSMC were plated at near confluence on glass chamber slides and then transfected overnight with 600 ng of siRNA to knockdown CCN5 levels or transfected with control siRNA. Following transfection VSMC were growth arrested by serum starvation for 72 hours. The confluent monolayers were then scratched uniformly with a pipette tip, excess cells were washed away, and serum free growth medium was added back. Initial pictures of the scratches were taken for reference (demarcation of initial wounds is represented by the white lines in each panel). The monolayers were then incubated for 48 hours at 37°C, fixed, stained with Diff-Quik to visualize cells and the extent of cell movement into the wound was measured by counting nuclei. The numbers below the panels represent the number of cells that have migrated into the wound area.