Induction by C2GnT of expression of the 130-kd CD43 glycoform in CD43 transfectant cells.

BALB/3T3 (3T3) fibroblastic cell lines (A,B) were transfected with CD43 cDNA (C,D) or cotransfected with CD43 and C2GnT cDNA (E,F). Stable transfectant cell lines were stained with S7 or MFT3 mAb followed by FITC-MAR18.5 and analyzed on a FACScan. The shaded histogram shows results from staining with the first mAb; and the open histogram, results from staining without the first mAb. Lysate from transfectant cell lines was subjected to SDS-PAGE (6% polyacrylamide gels), and Western blotting was performed with MFT3 mAb or FY14 mAb (IgG2a). The protein signals were detected by alkaline phosphatase–conjugated goat antirat IgG followed by 5-bromo-4-chloro-3-indolyl phosphate/nitro blue tetrazolium reagent (G) or by peroxidase-conjugated goat antirat immunoglobulin followed by enhanced chemiluminescence reagent (H).