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. 2019 Jul 30;47(19):e114. doi: 10.1093/nar/gkz659

Figure 2.

Figure 2.

Transduction pathway and integration competence of the RGD-modified lentiviral vector. (A) Subcellular localization analysis of Y77-RGD vector (red) by immunofluorescence labeling in HeLa cells. Scale bar, 20 μm. (B) Co-localization analysis of Y77-RGD (green) and Rab5/7-red fluorescent protein (RFP) (red) as markers of early/late endosomes, or Y77-RGD (red) and Golgi (green) in HeLa cells 6 h after viral transduction. Nuclei were stained with DAPI (blue). Scale bar, 20 μm. (C) Integration capability of the RGD-modified lentiviral vector analyzed by PCR amplification of GFP sequences using genomic DNA as templates. The genomic DNA was extracted from 293T cells transduced with wild-type or D192/Y77/Y116 ± RGD vectors. Genomic DNA extracted from blank 293T cells and a template using water or lentivirus were used as negative controls. PCR products were evaluated on a 1% agarose gel stained with GelRed. Marker, 1-kb ladder. (D) Quantitative analysis of the effects of cRGD modifications on viral transduction ability in 293T cells. The luciferase activity was tested 2, 3, 5, 7 and 9 days after transduction (***P < 0.001, n = 3).