Hiv-1 exploits ccr5 conformational heterogeneity to escape inhibition by chemokines

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Fig. S1. Native chemokines show weak anti-HIV-1 activities. CD4+ T-lymphocytes (A) or HeLa P4C5 cells (B) were inoculated with Bx08Ren viruses in the presence or in the absence of increasing concentrations of the indicated native chemokines or chemokine analogs. Results are expressed as percent infection relative to control infection measured in the absence of chemokines (100 %) and were fitted to a sigmoidal dose-response model with a variable slope. Representative experiments out of at least 3 independent experiments performed in triplicate are shown.

Fig. S2. PTX treatment of activated CD4+ T-cells (A-C) and HeLa P4C5 (D-F) cells neither influences HIV-1 infectivity nor the anti-HIV-1 activity of chemokines. Cells treated (C and F) or not (B and E) by PTX were infected as in Fig. S1 using 10 ng of p24 of the Bx08Ren, JRRen, 25Ren, 34Ren, 50Ren or 58Ren viruses in the absence (A and D) or in the presence of 10 nM CCL4 or 5P12-RANTES (B-C and E-F). Panels A and D are representative experiments. Panels B, C, E and F represent means ± SEM of four independent experiments performed in triplicate. For experiments in CD4+ T-cells, cells obtained from two different donors were used.

Fig. S3. G-protein dependent high-affinity binding of agonist chemokines to CCR5 is dispensable for chemokine-mediated inhibition of infection. PTX-treatment impaired specific binding of 0.1 nM 125I-CCL3 to HeLa P4C5 cells (A) but not the ability of chemokines to inhibit infection of these cells (B). In Panel A, results represent means ± SEM of eight independent experiments (***, p < 0.001 as compared to the untreated control in unpaired two-tailed Student’s t test). The other panel shows representative experiments out of at least three independent experiments.

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