Fig. 1

Knockout of FUT8 suppresses the invasive ability of two highly metastatic breast carcinoma cell lines. a and b Establishment of FUT8-knockout (KO) cells by CRISPR-Cas9-mediated genome editing. Protein domain organization of FTU8 is depicted on the top of each panel. TM, transmembrane domain. Two independent CRISPR-Cas9 clones targeting exon 3 or 6 of FUT8 were established (KO#1 or #2) in two invasive breast cancer cells, MDA-MB-231 (a) and Hs578T (b). Insertion (+) or deletion (Δ) mutations of each clone were verified by sequencing and are shown in parentheses. Inactivation of FUT8 gene and consequent loss of core fucosylation were validated by western blot analysis (lower-left panel) and core-specific Lens culinaris agglutinin (LCA) binding assay (lower-right panel). kDa, kiloDalton. Cell migration (c, d) and invasiveness (e, f) of control and FUT8-KO MDA-MB-231 (c, e) and Hs578T cells (d, f) were measured by Transwell assay with (invasion assay) and without Matrigel coating (migration assay). Data are mean ± SD. **p < 0.01, FUT8-KO versus parental cells. OD optical density