Figure 5.

Insertion of the IS2 Element or a Control 1.2-kb Fragment into the IDLVs Reduce the Total Number of Episomes and the Formation of Lower Expressing 2-LTR Circles

(A) Top, schematic representation of SE, SE-IS2, and SE-1.2kb eGFP; SFFV (spleen focus-forming virus) promoter. Bottom, representative plots showing eGFP expression profiles of 293T cells transduced with the different IDLVs at MOI = 0.7. The percentages (%) and expression levels (MFI) of the eGFP⁺ population are shown in each plot. (B) Graph showing the titer of the different IDLV supernatants using the ABM’s Lentiviral qPCR Titer Kit (see Materials and Methods for details). (C) Relative amounts of vector genomes in 293T cells transduced with the different IDLVs at 72 hr post-transduction and normalized to the levels observed in SE IDLVs (see Materials and Methods for details). (D) Graph showing the percentage of GFP positive cells in 293T cells transduced with the different IDLVs and normalized to the levels observed in SE-IDLVs. (E) Graph showing the relative amounts of 2-LTR circles relative to total viral genomes in 293T target cells at 72 hr post-transduction and normalized to the levels observed in SE-IDLVs (see Materials and Methods for details). The values represent means ± SEM of at least four separate experiments (*p < 0.05). The data were reported to an equal amount of cells, estimated by β-albumin housekeeping gene.