Figure 9. Anti-CD36 antibody treatment induces HIV-1 tethering at the plasma membrane of HeLa cells expressing CD36. Quantification of p24 Gag in the supernatant (A) and lysates (B) of HeLa and HeLa-CD36 cells treated with the FA6-152 mAb or with the IgG1 control overnight. A representative experiment is shown. The data shown are the means and SEM of triplicates. (C) Confocal sections of HeLa and HeLa-CD36 cells infected with HIV-1 Gag-iGFP, treated with the anti-CD36 mAb or its isotype control (Iso-Ab) for 48 h, and stained for CD9. Insets are enlargements of the boxed regions. Bars, 10 µm. Merge images include DAPI staining in blue. (D) Electron micrographs of HIV-1 NLAD8-infected HeLa-CD36 cells treated with FA6-152 mAb or its isotype control. Bar, 10 µm. (E) Multiple alignment illustrating the conserved residues present in the cytoplasmic tails of the proteins indicated. The figure was adapted from Primo et al. (2005). (F) Table showing the CD36 mutants produced with the corresponding changes in the C-tail sequence. The mutated amino acids are shown in red. The CD36467* and the CD36Y463F mutants are impaired for binding and capture of MOxLDL (Malaud et al., 2002) and for Staphylococcus aureus internalization (Stuart et al., 2005), respectively, whereas the CD36C464S mutant is impaired for S. aureus internalization (Stuart et al., 2005) and for loss of TSP-1–dependent VEGF-A165–induced migration (Primo et al., 2005). (G) Quantification of p24 Gag in the supernatant of HeLa cell lines stably expressing the indicated CD36 mutants, treated with the anti-CD36 mAb or its isotype control. CD36 cell surface expression was assessed by FACS (indicated by + or –). Data are presented as the means and SEM of three independent experiments performed on three different preparations of transduced cell lines, with the exception of the experiments with the 467* and C464S mutants which were independently repeated twice. Statistical analysis was performed by two-way ANOVA with Bonferroni’s post hoc test (*, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001).
Figure 9.

Anti-CD36 antibody treatment induces HIV-1 tethering at the plasma membrane of HeLa cells expressing CD36. Quantification of p24 Gag in the supernatant (A) and lysates (B) of HeLa and HeLa-CD36 cells treated with the FA6-152 mAb or with the IgG1 control overnight. A representative experiment is shown. The data shown are the means and SEM of triplicates. (C) Confocal sections of HeLa and HeLa-CD36 cells infected with HIV-1 Gag-iGFP, treated with the anti-CD36 mAb or its isotype control (Iso-Ab) for 48 h, and stained for CD9. Insets are enlargements of the boxed regions. Bars, 10 µm. Merge images include DAPI staining in blue. (D) Electron micrographs of HIV-1 NLAD8-infected HeLa-CD36 cells treated with FA6-152 mAb or its isotype control. Bar, 10 µm. (E) Multiple alignment illustrating the conserved residues present in the cytoplasmic tails of the proteins indicated. The figure was adapted from Primo et al. (2005). (F) Table showing the CD36 mutants produced with the corresponding changes in the C-tail sequence. The mutated amino acids are shown in red. The CD36467* and the CD36Y463F mutants are impaired for binding and capture of MOxLDL (Malaud et al., 2002) and for Staphylococcus aureus internalization (Stuart et al., 2005), respectively, whereas the CD36C464S mutant is impaired for S. aureus internalization (Stuart et al., 2005) and for loss of TSP-1–dependent VEGF-A165–induced migration (Primo et al., 2005). (G) Quantification of p24 Gag in the supernatant of HeLa cell lines stably expressing the indicated CD36 mutants, treated with the anti-CD36 mAb or its isotype control. CD36 cell surface expression was assessed by FACS (indicated by + or –). Data are presented as the means and SEM of three independent experiments performed on three different preparations of transduced cell lines, with the exception of the experiments with the 467* and C464S mutants which were independently repeated twice. Statistical analysis was performed by two-way ANOVA with Bonferroni’s post hoc test (*, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001).

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