Mzfliii system

Cells were plated in 12-well plates on both sides of a plexiglass insert that was removed once the cells had reached confluence (T0). Cells in the entire well were imaged at T0 and T16 (T = 16 h) and the wound closure was calculated by the difference of the covered area by the cell monolayer between T0 and T16. Images were taken with a Leica MZFLIII system through an Axiocam HRc from Zeiss. Results are mean of three independent experiments performed in triplicate.
Videomicroscopy of migrating HaCaT cells on a fibronectin matrix was performed on a Biostation (Nikon) during 24 h after PDMS insert removal. Cell trajectories were manually obtained using the MTrackJ ImageJ plugin. Cell speed was calculated by a regression analysis of the plot representing the distance versus the time. Directionality was calculated as cosθ where θ is the angle between the field vector at t = 18 h and the cell migration direction; the persistence was calculated after 18 h as the ratio of the cumulative distance over the Euclidian distance. Velocity correlation length was measured as described previously^{15 (link)}. The separation time was estimated manually by following two cells that were neighbours at t = 0 h and measuring the time required for the cells to separate from each other.

Cells were plated in 12-well plates on both sides of a plexiglass insert that was removed once the cells had reached confluence (T0) and cell culture medium was replaced by serum-free medium. Cells in the entire well were imaged at T0 and T16 (T = 16 h) and the wound closure was calculated by the difference of the covered area by the cell monolayer between T0 and T16. Images were taken with a Leica MZFLIII system through an Axiocam HRc from Zeiss. Results are mean of three independent experiments performed in triplicate.