Fig. 2

Fucosyltransferase 8 (FUT8) plays a critical role in transforming growth factor-β1 (TGF-β1)-induced epithelial–mesenchymal transition (EMT). a Knockdown of FUT8 expression in MCF-10A cells. MCF-10A cells were infected with a lentiviral vector to generate a stable clone expressing a short hairpin RNA (shRNA) control or two independent shRNAs targeting FUT8 (FUT8-shRNA #1 and #2), respectively. The efficiency of FUT8 knockdown in the selected stable pool was confirmed by western blot analysis. α-Tubulin expression was an internal control. b FUT8 knockdown impaired the EMT in MCF-10A cells. Control or FUT8-knockdown MCF-10A cells were treated with TGF-β1 (10 ng/ml) for 8 days. The effect of FUT8 knockdown on the EMT was examined by measuring E-cadherin and vimentin expression by western blot analysis as previously described. c Overexpression of FUT8 produces invasive, nonpolar, and disorganized acini in MCF-10A cells. MCF-10A cells were infected with a lentiviral vector to generate a stable clone expressing control (pSin) or FUT8 (pSIN-FUT8). FUT8 overexpression was confirmed by western blot analysis (left panel). FUT8-overexpressing MCF-10A cells show invasive and disorganized acini on 3D culture (right panel). d Overexpression of FUT8 promoted EMT-like phenotype in MCF-10A cells. Control or FUT8-overexpressing MCF-10A cells were treated with TGF-β1 for 8 days. The EMT was determined by measuring the E-cadherin and vimentin expression (top panel). FUT8 expression was examined by western blot analysis (bottom panel). α-Tubulin expression was examined as internal control individually. kDa, kiloDalton