Figure 6.

CD81 tetraspanin surface intensity and viral production. HeLa cells and HeLa cells stably expressing Myr (+) Ank^GAG1D4-GFP or Myr (+) Ank^A32D3-GFP were transduced with VSV-G-pseudotyped HIV-1 GagmCherry. (A) After 48 h, the level of ankyrin expression and HIV-1 GagmCherry expression was analysed by flow cytometry. Populations were defined based on their fluorescence intensity ((+) = intermediate expression, (++) = high expression). (B) Surface CD81 expression on non-transduced cells was measured by flow cytometry. The intensity of CD81 on non-transduced HeLa and ankyrin-expressing HeLa cells was shown in Bar graph. (C) Surface CD81 expression on transduced HeLa and ankyrin-expressing HeLa cells was determined by flow cytometry. Relative surface CD81 intensity was calculated by the percentage of surface CD81 intensity of transduced cells and normalised against surface CD81 intensity of non-transduced cells. Data represent the mean ± SD from triplicate independent assay. Significant differences (*** p < 0.001; ns, not significant) were determined between transduced and non-transduced HeLa cells or ankyrin-expressing HeLa cells using one-way ANOVA. Ank^GAG1D4, Ank^A32D3 represent HeLa cells stably expressing Myr (+) Ank^GAG1D4-EGFP and Myr (+) Ank^A32D3-EGFP, respectively.