Fig 4.

Binding of GT24-Dynabeads to heparan sulfate on the cell surface. (A) Pulldown of cells using GT24-Dynabeads. Biotinylated GT24 peptide was immobilized onto streptavidin-coupled Dynabeads. GT24-Dynabeads were incubated with cells for 30 min at room temperature and separated from supernatant using a magnet. Binding assays for U251 and Jurkat cells were carried out in 1× PBS, pH 7.4, while binding assays for Sf9 insect cells were performed in insect cell culture medium with a pH of 6.2. Bead-bound cells were resuspended in the same volume of the supernatant and counted on a hemocytometer. Images of cells before separation and those remaining in the supernatant were also taken. (B) Transduction of Jurkat T cells with CMV-EGFP and VSVG-CMV-EGFP baculovirus. VSVG-CMV-EGFP baculovirus expresses both VSVG and the endogenous gp64 on the virus surface while CMV-EGFP virus has only gp64. One day after transduction, the EGFP-positive Jurkat cells were analyzed using flow cytometry.