Fig. 7. Lack of focal adhesions resulting from PKC inhibition can be overcome by LPA treatment but not by HepII and results in recruitment of syndecan-4. (A) REF cells were plated on FN110-coated coverslips for 1.5 hours and either treated with DMSO or with 0.5 µM Gö6976. Cells were subsequently cultured without further treatment (–) or treated with LPA, recombinant HepII, or vehicle as indicated in the Materials and Methods for 30 minutes and cells were fixed, permeabilized and double stained for F-actin (red) and vinculin (green). The percentage of cells forming stress fibres and focal adhesions under the various treatments is indicated. Results are representative of at least three independent experiments. Bar, 50 µm. (B) REFs were plated on FN- or FN110-coated coverslips. At 2 hours, cells on FN110 were either left untreated (–) or treated with LPA, HepII, or pre-treated with Gö6976 for 30 minutes before LPA stimulation, as described above. Cells were subsequently fixed and stained for syndecan-4 and -actinin. Arrows indicate localization of syndecan-4 and -actinin to focal adhesions, arrowheads to focal complexes. Bar, 50 µm.