Fig. 3

Surface SSEA-1 level was positively correlated with B3GNT5 expression. A The schematic diagram of SSEA-1 antigen formation. After the initiation of GSLs biosynthesis from ceramide (Cer) to glucosylceramide, B3GNT5 is responsible for Lc3Cer formation. Elongation of Lc3Cer by β-1,4-galactosyltransferase 4 (B4GALT4) and α-1,3-fucosyltransferase 4 (FUT4) yields the SSEA-1 antigen. B Expression of B3GNT5 was examined by Western blotting in MDA-MB231 and BT549 cells with stable empty vector or B3GNT5-WT-Flag cDNA. C, D Surface expression of SSEA-1 antigen was tested by flow cytometry in MDA-MB231 (C) and BT549 cells (D) with stable empty vector or B3GNT5-WT-Flag cDNA. E Expression efficiency of B3GNT5 was analyzed by Western blotting in wild-type MDA-MB231 and BT549 cells (WT) as well as B3GNT5-knockout MDA-MB231 and BT549 cells with stable empty vector (KO) and B3GNT5-WT-Flag cDNA (KO + B3GNT5). F, G Flow cytometry measuring surface SSEA-1 level in the indicated cell lines. Data are presented as the percentage of wild-type vector cell line. *p < 0.05 by Student’s t test. Data are shown as mean ± SD based on three independent experiments