Figure 2

Transduction of c-kit-expressing cells using lentiviral vectors bearing a VSV-G-derived fusion domain and SCF. (A) Schematic representation of the VSV-GS fusion protein. HA: An HA epitope sequence (KYPYDVPDYA) was included to facilitate detection of the VSV-GS protein. The truncated ectodomain, the transmembrane domain (TM) and the cytoplasmic tail (CT) are indicated. The numbers refer to the ends of the respective protein domains. (B) Transduction of 293T and 293-c-kit cells using LV-EGFP vector particles bearing the VSV-GS fusion domain and/or SCF. Additional controls included LV-EGFP vector particles bearing VSV-GS but lacking SCF and vector particles lacking both VSV-GS and SCF. Left panels: 293T cells; Right panels: 293-c-kit cells. Cells were analyzed by FACS three days later. FACS profiles of representative assays are shown. (C) Transduction of MO7-e cells using EGFP-encoding lentiviral vector particles displaying SCF plus the VSV-GS fusion domain. Cells were transduced by spinoculation. Vector titers were determined by FACS analysis three days later. Goat anti-hSCF was added during transduction of the samples shown in the lower center and lower right panels. Normal goat serum referred to as control serum was added to the sample shown in the bottom left panel.