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Figure 8 | BMC Biotechnology

Figure 8

From: Generation and characterization of a stable cell population releasing fluorescent HIV-1-based Virus Like Particles in an inducible way

Figure 8

Efficient delivery of the NefG3C-GFP-related fluorescence by pseudotyped 18-4s VLPs. (A) Western blot analysis of NefG3C-GFP pseudotyped VLPs. Hundred ng of differently pseudotyped VLPs were analyzed by Western blot for the presence of NefG3C-GFP and of the respective receptors. The migration of major viral products is indicated on the left side, whereas the molecular marker sizes are reported on the right. (B) FACs analysis of CEMss cells 2 and 4 hours after the challenge with 250 ng/105 cells of NefG3C-GFP VLPs pseudotyped with VSV-G or, as control, with its mutant defective for the fusion activity (fm). As control, in a separate experiment the same amounts of (VSV-G) VLPs were used to challenge cells untreated (Ctrl) or pre-treated for 2 hour at 37°C with either 100 nM bafilomycin A1 or 50 μM chloroquine. Cells were analyzed 2 hours after the challenge. Bars were drawn at the highest fluorescence levels of untreated cells. Percentages of GFP positive cells are reported in the respective plots. (C) FACs analysis of CEMss cells 2 and 4 hours after the challenge with 1 μg/105 cells of NefG3C-GFP VLPs pseudotyped with X4-tropic HIV-1 Env or, as control, without receptors ("Null"). As control, in a separate experiment the same amounts of HIV-1 Env VLPs were incubated for 1 hour at 4°C with the 17b anti-Env gp120 neutralizing mAb, or with an isotype matched irrelevant mAb (Ctrl) before the cell challenge. Cells were analyzed 4 hours after the challenge. Bars were drawn at the highest fluorescence levels of untreated cells. Percentages of GFP positive cells are reported in the respective plots.

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