Figure 5.

A, B) H358 cells were cultured on 40-mm Bioptechs coverslips before wounding with a single scratch and culturing under control conditions (RPMI plus 5% charcoal-stripped FBS; A) or in the presence of TGF-β1 (2 ng/ml; B) for up to 48 h. Time-lapse phase microscopy was performed using a Nikon TE2000; culture medium was saturated with a gas mixture containing 5% CO₂ and 21% O₂, perfused through an environmental control system chamber. Images were collected with a Cascade EMCCD camera with on-chip multiplication gain driven by MetaMorph software. Images were acquired every 60 s, with an exposure time of 32 ms. Representative photomicrographs are shown at ∼30 h postwounding after tracking the positions of cells for 6 h. Insets: higher-magnification views of cells. Asterisks indicate cells that were tracked and shown in the graphs in C, D. C, D) Rose plots show individual cell velocities in the x and y directions. Colored lines correspond to individual cells marked by asterisks in A, B. E) Graph represents means ± sd of 6 cells tracked in 2 independent determinations. **P < 0.005.