Receptor-Targeted Nipah Virus Glycoproteins Improve Cell-Type Selective Gene Delivery and Reveal a Preference for Membrane-Proximal Cell Attachment
Fig 9
CD117- and GluA4-targeted NiV-LVs.
(A) Surface expression of CD117 (blue line) and CD117short (red line) on stably expressing HT1080 cells compared to the parental cell line (filled curve) as determined by flow cytometry. Cells were stained with PE-coupled CD117 antibody. One representative out of four experiments is shown. See S10 Fig for quantitative data. (B) Surface expression of GluA4 (blue line) and GluA4short (red line) on stably expressing HT1080 cells compared to the parental cell line (filled curve) as determined by flow cytometry. Cells were stained with PE-coupled myc-tag antibody. One representative out of three experiments is shown. See S10 Fig for quantitative data. (C) Binding of recombinant SCF to CD117 and CD117short. Fc-SCF was produced in HEK-293T cells by transient transfection. HT1080, HT1080-CD117 and HT1080-CD117short were incubated for 1 h at 4°C with the same volumes of recombinant Fc-SCF prior to staining against Fc-tag using FITC coupled anti-Fc antibody. One representative out of three experiments is shown. See S10 Fig for quantitative data. (D) Titers of concentrated stocks of NiVmutCD117-LV and NiVmutGluA4-LV. NiVmutCD117-LV was titrated on HT1080-CD117 and HT1080-CD117short cells (n = 4; mean ± standard deviations (SD) are shown; **, P<0.01by unpaired t-test). NiVmutGluA4-LV was titrated on HT1080-GluA4 and HT1080-GluA4short cells (n = 3; mean ± standard deviations (SD) are shown; ***, P<0.001 by unpaired t-test).