Figure 8.

ES6A filopodia and lamellipodia are blocked by anti-α6 antibody GoH3 and are restored by cross-linking GoH3 with secondary antibody. Glass coverslips were coated for 30 min at 37°C with 40 μg/ml Fn. ES6A, ES6B, or ESneo cells (1 × 10⁴ cells/coverslip) were plated. For antibody treatments, cells were incubated at room temperature with 10 μg/ml GoH3 or 10 μg/ml goat anti-rat affinity-pure antibody in migration medium for 15 min. For cross-linking, 10 μg/ml goat anti-rat affinity-pure antibody was added for an additional 15 min to cells that had bound GoH3. No antibodies were removed before plating on the coverslips. Cells were maintained at 37°C for 18 h, then washed with PBS, fixed in 2.5% paraformaldehyde in PBS, and mounted in Immunofluore. Cells were visualized in phase contrast by using a Zeiss Axiovert microscope and photographed using TMAX 400 ASA film. (A, D, and G) ES6A cells: control, GoH3, and cross-linked GoH3, respectively. (B, E, and H) ES6B cells: control, GoH3, and cross-linked GoH3, respectively. (C, F, and I) ESneo cells: control, GoH3, and cross-linked GoH3, respectively. Bar, 20 μm.