Adhesive wound assay inserts

Adhesive wound assay inserts (Ibidi) were used to generate wounds in confluent layers in a 6 well plate. To do this, transfected cells were trypsinized and seeded into the plate to form a monolayer. After overnight growth and attachment, the insert was carefully removed and the cells were starved of serum by adding serum free media for 4 hours, after which DMEM was added to the cells and they were imaged immediately (T0) using (Olympus cellSens Dimension 1.12). Cells were maintained in an incubator at 37°C, 5% CO₂ for 24 hours, after which time images of the wound closure were taken as described above. The percentage wound closure was analysed using Wimasis image analysis.

Adhesive wound assay inserts (Ibidi) were used to generate wounds in confluent layers in a 6 well plate. To do this, stable cells were trypsinized and seeded into the plate to form a monolayer. After overnight growth and attachment, the insert was carefully removed and the cells were starved of serum by adding serum free media for 4 hours, after which DMEM was added to the cells and they were imaged immediately (T0) using (Olympus cellSens Dimension 1.12). Cells were maintained in an incubator at 37^°C, 5% CO₂ for 24 hours, after which time images of the wound closure were taken as described above. The percentage wound closure was analyzed using Wimasis image analysis.