2002
DOI: 10.1074/jbc.m208283200
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The α(1,3)-Fucosyltransferase Fuc-TIV, but Not Fuc-TVII, Generates Sialyl Lewis X-like Epitopes Preferentially on Glycolipids

Abstract: Fuc-TIV and Fuc-TVII are the two ␣(1, 3)-fucosyltransferases in myeloid cells responsible for the biosynthesis of sialyl Lewis X (sLe x ), the minimal ligand structure for the selectins. We have compared the ability of Fuc-TIV and Fuc-TVII to generate sLe x -like epitopes in transfected Chinese hamster ovary (CHO)-Pro ؊ 5 cells expressing the P-selectin glycoprotein ligand-1 and the core-2 branching enzyme C2GnT. We found that mouse Fuc-TIV and Fuc-TVII can generate similar levels of cell surface sLe x . Surpr… Show more

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Cited by 40 publications
(31 citation statements)
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References 54 publications
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“…26 For cDNA synthesis, 2 g of total RNA was used as template in a 25-L reverse transcription reaction. PCR reactions were incubated for 5 minutes at 95°C, followed by 35 amplification cycles with 30 seconds of denaturation at 94°C, 30 seconds of annealing at 60°C, and 30 seconds of extension at 72°C.…”
Section: Rt-pcr and Real-time Pcrmentioning
confidence: 99%
“…26 For cDNA synthesis, 2 g of total RNA was used as template in a 25-L reverse transcription reaction. PCR reactions were incubated for 5 minutes at 95°C, followed by 35 amplification cycles with 30 seconds of denaturation at 94°C, 30 seconds of annealing at 60°C, and 30 seconds of extension at 72°C.…”
Section: Rt-pcr and Real-time Pcrmentioning
confidence: 99%
“…Alternatively, matched cDNA pairs of colon cancer patients (BD Biosciences Clontech) were used for PCR. To reflect the initial mRNA expression levels, PCR amplification of cDNA was carried out with cycle numbers that have been tested to be in the linear range, as previously described (Huang et al, 2002). For b-actin detection, the sense and antisense primers were 5 0 -GCT CGT CGT CGA CAA CGG CT-3 0 and 5 0 -AAA CAT GAT CTG GGT CAT CTT CT-3 0 , respectively, generating a 326 base-pair fragment.…”
Section: Cancer Profiling Arraymentioning
confidence: 99%
“…Huang et al [48]. Single-cell suspensions of endothelial cell lines were obtained by incubating the monolayers with 0.05% Trypsin/0.02% EDTA solution (Biochrom, Berlin, Germany) whereas transfected CHO and 293T cells were harvested with 5 mM EDTA in PBS.…”
Section: Flow Cytometrymentioning
confidence: 99%
“…For immunoprecipitation, cells were surfacebiotinylated as described [49], lysed, and immunoprecipitated with 10 lg protein A-Sepharose-bound antibody according to [48]. For each immunoprecipitation CHO lysate containing 0.7 mg protein or DC lysate containing 2.1 mg protein was used.…”
Section: Western Blot Analysis and Immunoprecipitationmentioning
confidence: 99%