FIG 8.
Impact of mutations at Vpu E59/L63/V64 and S52/S56 residues on the sum of the VCC volume. MDMs were infected with VSV-G-pseudotyped WT, Udel, ELV, or 2.6 NL4-3 HIV-1. (A) At 5 days postinfection, cells were fixed and immunolabeled for BST2, CD81, and CAp24. The volumetric analysis of VCCs was performed using Imaris 7.2 software (Bitplane) by 3D reconstruction of MDMs. An example of 3D reconstruction of MDMs is shown. The merge presented on the right is the overlap of the three two-dimensional (2D) images corresponding to the 3D panels shown. Arrows show structures considered VCCs. (B and C) Histograms showing the sum of the VCC volume per cell obtained by thresholding fluorescence intensity values on 62 cells under each condition from three independent donors. Statistical analysis was done using one-way ANOVA combined with Bonferroni’s post hoc test (means ± SEM; n = 3 donors). ***, P ≤ 0.001; *, P ≤ 0.05. Bars, 20 μm.