Figure 1. Lentiviral vector and transduction procedures.

(A) Schematic diagram of Cell Genesys SIN lentiviral transfer vector pKC.MND.eGFP.neo. CMV 5′ LTR: modified cytomegalovirus 5′ long terminal repeat; RRE: rev response element; cPPT/CTS: central polypurine tract/central termination sequence; MND: myeloid proliferative sarcoma virus promoter; eGFP: enhanced green fluorescent protein; SV40: simian virus 40 promoter; neo: neomycin resistance gene; WPRE: woodchuck post translational regulatory element; ΔU3 LTR: modified U3 long terminal repeat. (B) Schematic diagram of experimental procedures for CD34⁺ transduction. Cells were pre-stimulated for 48, 24 or 4 h prior to transduction at T₀ at various MOIs, with and without retronectin and/or spinoculation. Virus was washed from the cells 2, 6, 24 or 48 h post transduction, placed back into culture and analysed by flow cytometry 48 h post transduction.