Figure 3.

Expression of Galα1,3Gal residues at the surface of transduced cells. Cells stably transduced or not with CMV promoter (pCMV)/murine α1,3galactosyltransferase (mαGalT), pNeu664/mαGalT, or pNeu250/mαGalT were labeled with fluorescein isothiocyanate-conjugated Griffonia simplicifolia/isolectin B4 (FITC-GS-I-B4) lectin (Galα1,3Gal) and analyzed on a FACScan cytometer (counts, cells numbers; FL1-H, fluorescence intensity). (a) Binding of FITC-GS-I-B4 to parental and pNeu250/mαGalT transduced HEK-293, MCF-7 and SK-BR-3 cells. Mean values of fluorescence detected by flow cytometry at the surface of stably transduced cells with (b) the constructs pNeu664/mαGalT or pNeu250/mαGalT in a plasmid vector (experiments in triplicates), and (c) the construct pNeu250/mαGalT in a self-inactivating retroviral vector (experiments were conducted using four to five subclones).