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. 2018 Jun 4;215(6):1679–1692. doi: 10.1084/jem.20172048

Figure 3.

Figure 3.

UGT8 activates the sulfatide biosynthetic pathway and enhances breast cancer cell migration and invasion. (A) Sulfatide biosynthetic pathway. (B) Stable transfectants with empty vector or knockdown of UGT8 expression were established in MDA-MB231 and SUM159 cells, and stable clones with empty vector or UGT8 expression were also generated in BT549 and HCC1937 cells. (C) Expression of GalCer and sulfatide was examined by immunoblotting in MDA-MB231 cells with stable empty vector or knockdown of UGT8 expression, as well as HCC1937 cells with stable empty vector or UGT8 expression. (D) Expression of GalCer and sulfatide was measured by immunofluorescent staining in MDA-MB231 cells with stable empty vector or knockdown of UGT8 expression as well as BT549 cells with stable empty vector or UGT8 expression. Nuclei were visualized with DAPI (blue). Bars, 20 µm. (E and F) Migratory ability (E) and invasiveness (F) of BT549 cells with stable empty vector or UGT8 expression as well as BT549 cells treated with or without GalCer (2 µM) or sulfatide (2 µM) were analyzed. The percentage of migratory and invasive cells was shown in the bar graph (mean ± SD in three separate experiments). (G and H) Migratory ability (G) and invasiveness (H) of MDA-MB231 cells with stable empty vector or knockdown of UGT8 expression, as well as shUGT8-expressing MDA-MB231 cells treated with or without GalCer (2 µM) or sulfatide (2 µM) were analyzed. The percentage of migratory and invasive cells was analyzed as in E and F. *, P < 0.01 by Student’s t test. Data are shown as mean ± SD based on three independent experiments.