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. 2016 Feb 1;97(Pt 2):422–434. doi: 10.1099/jgv.0.000341

Fig. 3.

Fig. 3.

HSV infection results in increased HPV16 internalization and percentage of cells with reporter gene transduction restricted to non-HSV-infected cells. (a) HaCaT cells were mock or HSV-1 infected for 2 h. Inoculum was removed, media replaced and cells incubated for 24 h. pHrodo Red-labelled HPV16 L1L2 VLPs were then added for 12 h. VLP internalization was assessed by flow cytometry and results are displayed as mean ± sd percentage positive cells of triplicate wells normalized to the mock-infected groups. H16.E70 was used as a positive control for blocking internalization. Data are representative of two independent experiments. (b) HaCaT cells were mock or HSV-2 infected as described above and incubated for 48 h. pHrodo Red-labelled HPV16 L1L2 VLPs were then added to the cells for 12 h. VLP internalization was assessed by flow cytometry and results are displayed as mean ± sd mean fluorescence intensity (MFI) of triplicate wells. Results are a representative example of two independent experiments. (c) Flow plot data showing HPV16 VLP uptake (x-axis) and HSV-2 infection (y-axis). Results are a representative example of two independent experiments. (d) HaCaT cells were infected with HSV-1 as described above. HPV16 PsVs containing a GFP reporter plasmid were then added and GFP expression was analysed 48 h post-infection by flow cytometry. Results are presented as mean ± sd infection of triplicate wells normalized to mock-infected groups. Data are representative of three independent experiments. (e) HaCaT cells were infected with HSV-2 as described above and incubated for 48 h. HPV16 WT PsVs containing a GFP reporter plasmid were then added to the cells and GFP expression was analysed 48 h post-infection by flow cytometry. Results are presented as mean ± sd gene transduction of triplicate wells. Data are representative of three independent experiments. (f) Flow plot data showing percentage of cells with HPV16 PsV gene transduction (y-axis) and with detectable HSV-2 infection (x-axis). Results are a representative example of two independent experiments. **P < 0.01 as determined by a one-way ANOVA followed by Tukey's multiple comparison test against the mock-treated group.