Fig. 3: ERVI impairs HIV-1 entry but not binding to TZM-bl target cells.

a Experimental workflow. HIV-1 NL4.3 Vpr.mRuby2 virions were cultured in S, DC or LC for 16 h. Equivalent amounts of RT units from culture supernatants or untreated virions were then incubated with TZM-bl cells for 2 h at 4 °C. Cell membranes were then stained with Concanavalin-A AF-488, prior to microscopy processing. b Normalized infectivity of HIV-1 NL4.3 Vpr.mRuby2 virions after 16 h of culture as in a. c Representative spinning disk micrographs of cells incubated with HIV-1 NL4.3 Vpr.mRuby2 virions. Yellow arrows indicate Vpr.mRuby2+ spots detected at the surface of the target cells. Scale bar: 15 µm. d Average binding frequency of HIV-1 NL4.3 Vpr.mRuby2 virions to target cells as shown in c. e Average volume of Vpr.mRuby2 spots detected as shown in c. f Experimental workflow. HIV1 NL4.3 Vpr.BlaM virions were as in a. Equivalent amounts of RT units from culture supernatants or untreated virions were then incubated with TZM-bl cells for 4 h at 37 °C, in the presence or absence of T20 fusion inhibitor. Cells were then loaded with the CCF2-AM and processed by flow cytometry. g Normalized infectivity of HIV-1 NL4.3 Vpr.BlaM virions after16h of culture as in a. h Representative flow cytometry dot plots of CCF2 loaded cells. Gates identify cleaved-CCF2+ cells. See Sup. Figure. 9a for gating strategy. i Quantification of the percentage of CCF2-product positive cells measured by flow cytometry. j Correlation between Relative Infectivity and levels of CCF2-product positive cells after ERVI by linear regression. Results represent the mean ± SD of three independent experiments. Symbols indicate individual experiments. Significance is indicated by p-values, and was calculated by one-way ANOVA test; Dunnets’s post-test (b, g), or by one-way ANOVA test; Tukey’s post-test (d, e, i). Source data are provided as a Source Data file.