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Fig. 4. The ability of ST6GalNAc I transfectants to migrate but not to adhere on fibronectin is impaired. (A) The motility of STn- and STn+ cells on various extracellular matrix components was quantified by image analysis in a phagokinetic track assay. The extracellular matrix components coated on wells of culture plates were BSA, fibronectin (FN), collagen (C) I, collagen IV, collagen VI and hyaluronic acid (HA). All were used at 10 µg/ml except for hyaluronic acid, which was used at 5 µg/ml. Error bars represent the standard error of three experiments. The differences in migration between STn+ and STn-cells on fibronectin and hyaluronic acid were significant (P<0.001 and P<0.05, respectively); the difference on collagen was not significant. (B) Representative fields of phagokinetic tracks on fibronectin from one STn- and one STn+ clone. (C) Inhibition by soluble fibronectin of the STn- and STn+ cells' motility on coated fibronectin in a phagokinetic track assay. Treated cells vs control cells: P<0.001. (D) Motility on fibronectin of the STn+ transfectant clones compared with that of the STn-control clones quantified by a phagokinetic track assay. Error bars represent the standard deviation of the mean from ten independent experiments. STn+ vs STn-: P<0.001. (E) Motility on fibronectin (10 µg/ml) of STn+ and STn-cells in a transwell assay was quantified by image analysis. Error bars represent the standard error of the mean of three experiments (P<0.001 between STn- and STn+ cells). (F) Adhesion on fibronectin coated at different concentrations of STn-(blue circles) and STn+ (red circles) cells measured at 15 minutes and 40 minutes after seeding. Error bars represent the standard deviation of the mean from eight values. In all experiments, the values obtained for the two STn-control clones (B10 and F4) and the two STn+ ST6GalNAc I transfectants (C2 and G9), respectively, were pooled.
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