Wound healing assay

For wound closure assay, 1.5 × 10⁴ cells in 100 µl media were seeded into each well of a 96-well ImageLock Microplate (Essen BioScience Cat #4379) and incubated overnight. Using a 96-pin IncuCyte woundmaker, the monolayer was gently scratched across the well and washed with medium to remove cell debris. Fresh medium was then added, cells were incubated and images of scratched wounds were captured at different time points using the IncuCyte ZOOM® live-cell system (Essen Bioscience). Results were analyzed using the IncuCyte software and presented as relative wound density.