The ectopic expression of FUK in melanoma cells inhibits invadopodium formation and delays invadopodial initiation.

<p><b>A,</b> the effect of FUK overexpression on the fucosylation of WM793 cell surface proteoglycans. The plasma membrane proteins from control and FUK OE WM793 cells were separated using SDS-PAGE and detected by biotin-UEA1. <b>B,</b> the effect of FUK overexpression on cell surface fucosylation as detected by UEA1 staining and flow cytometry. <b>C</b>, representative images showing the effects of ectopically expressed FUK on invadopodium formation and TexasRed gelatin degradation in WM793. <b>D and E,</b> quantitation of the effect of ectopically expressed FUK on the invadopodium number per cell (<b>D</b>) and the proportion of invadopodia positive cells (<b>E</b>) in WM793. <b>F,</b> quantitation of the effects of FUK overexpression on gelatin degradation. <b>G,</b> the effects of ectopic FUK on the initiation of the assembly of invadopodia induced by 10% FBS. <b>H,</b> qPCR assay showing the effects of FUK sh1+sh2 on the mRNA transcript levels of FUK in WM793 cells. <b>I,</b> quantitation of the effects of FUK knockdown on fucose-mediated inhibition of invadopodium formation in WM793 cells. *** indicates <i>p<0</i>.<i>001</i>, as determined by two-tailed, two sample t-test (<b>D, G, H, I</b>) or two-tailed Fisher’s exact test (<b>E</b>). Numbers in parentheses indicates the number of cells used in quantitation. Representative results from at least three independent replicates were presented.