CD2AP suppresses cell motility and wound-induced cell migration. (a) Time-lapse phase-contrast microscopy showing migration of CD2AP knockdown cells into free space, whereas parental MDCK cells are relatively stationary. (b) Time-lapse phase-contrast microscopy showing wound-induced migration of 2 d postconfluent MDCK cell monolayer. (c) Quantitation of migration speed in MDCK cells showing decreased migration rate as a function of time (P < 0.0001). Representative data from three separate experiments (n = 3) are shown. (d) Line tracings of wound edge at 1-h intervals showing a decrease in migration rates of parental but not CD2AP knockdown cells at 4 and 6 d postconfluency (4 d and 6 d). (e) Line tracings of wound edge at 1-h intervals showing suppression of wound-induced migration of cells expressing exogenous CD2AP at 1 and 2 d postconfluency. (f) Quantitation of migration speed in CD2AP knockdown cells showing a lack of suppression in wound-induced migration after confluency. Representative data from three separate experiments (n = 3) are shown. (g) Actin (phalloidin) staining showing correlation of cell migration with wound-edge protrusions (white arrowheads). Protrusions were suppressed in 4 d and 6 d postconfluent parental cells and cells expressing exogenous CD2AP. (h) Quantitation of migration speed showing suppression of wound-induced migration in 1 d and 2 d postconfluent cells expressing exogenous CD2AP (P < 0.0001). Representative data from three separate experiments (n = 3) are shown.