Figure 1
Antibody and H RBD binding following transfection of the Glut-1 and Glut-3 glucose transporters. (A) 293T cells were transfected with Glut-1 or Glut-3 expression vectors and assayed for binding to the mAb1418 and C-term anti-Glut-1 polyclonal Ab. The former stainings were performed on whole cells as well as permeabilized cells to determine cell surface and total binding, respectively. All stainings using the anti-Glut-1 pAb were performed on permeabilized cells as the recognized epitope is intracellular. Staining was performed at 4°C. Specific binding and background fluorescence due to the secondary conjugated Ab are indicated in solid line and filled histograms, respectively. (B) Control and transfected 293T cells were incubated with rFc-tagged H1RBD and H2RBD fusion proteins for 30 min at 37°C followed by incubation with a FITC-conjugated αrabbit-Fc antibody at 4°C. Direct binding to H2RBD was demonstrated by incubation of cells with an EGFP-tagged envelope (H2RBD-EGFP). Binding is shown in solid line histograms whereas control immunofluorescence is shown in filled histograms.