Mitomycin c

Cells were seeded into 2-well silicone insert (Ibidi, cat# 80209) at a density in which the cells are 100% confluent 24 h later. The next day, cells were treated with mitomycin C (MedKoo Biosciences, cat# 100630) at 10 μg/mL for 2 h. After 2 h, the insert was removed and the cells were washed with PBS 3 times before replacing with growth medium. The size of the wounds at this time (0 h), and at different time intervals, were monitored using either the EVOS cell imaging system or the Incucyte® S3 Live-Cell Analysis System. The percentage wound healing at the various time points (i.e., x h) was calculated based on the equation below: $\%\text{Wound}\text{healing}=\frac{\text{Size}\text{of}\text{wound}\text{at}0\mathrm{h}-\text{Size}\text{of}\text{wound}\text{at}x\mathrm{h}}{\text{Size}\text{of}\text{wound}\text{at}0\mathrm{h}}\times 100$