Fig. 5
From: FUT8 promotes breast cancer cell invasiveness by remodeling TGF-β receptor core fucosylation
Transforming growth factor-β (TGF-β) receptor proteins I and II (RI and RII) are core fucosylated by fucosyltransferase 8 (FUT8) in HEK-293 T cells. a Establishment of FUT8-knockout (KO) HEK-293 T cells by CRISPR-Cas9-mediated genome editing. Two independent CRISPR-Cas9 clones targeting exon 3 or 6 of FUT8 were established (upper panel). Inactivation of FUT8 gene and consequent loss of core fucosylation were validated by western blot (lower-left panel) and LCA binding assay (lower-right panel). b Protein domain organization of TGF-β RI and RII. According to the Uniprot database (http://www.uniprot.org), potential N-linked glycosylation sites within TGF-β RII (Asn-70, 94, and 154) and RI (Asn-45) were marked. SP, signal peptide sequence; TM, transmembrane domain. c Core fucosylation of TGF-β RI and RII proteins were eliminated by FUT8 knockout. Recombinant TGF-β RI or RII proteins in the control or two FUT8-KO 293 T cell lines were probed with biotinylated Lens culinaris lectin (LCA), then detection with streptavidin-conjugated horseradish peroxidase. kDa, kiloDalton