FIGURE 6.

Overexpression of ST6GAL1 was able to neutralize the anti-migratory role of GnT-III in HeLa cells. A, cell lysates from the control and GnT-III-overexpressing HeLa cells were immunoblotted with E4-PHA and DSA lectins to check the overexpression efficiency of GnT-III. B, these two cells together with ST6GAL1 and GnT-III-overexpressing HeLa cells were incubated with (bold line) or without (gray shading) biotin-conjugated MAA or biotin-conjugated SNA, followed by incubation with streptavidin Alexa Fluor 647 conjugate and subjected to FACS analysis. C, their cell migration toward FN was determined by Transwell assay. Cells that migrated through the Transwell membrane were stained with 0.5% crystal violet. Shown is a representative example recorded by phase contrast microscopy. Scale bar, 200 μm. The migrated cells were counted under a microscope. The quantitative data were obtained from three independent experiments. The p values were calculated using one-tail unpaired t test. Error bars indicate standard derivation. *, p < 0.01. ST6GAL1 OE, ST6GAL1-overexpressing cells; Con, control; OE, GnT-III-overexpressing cells.