Fig. 7: Vpr enhances HIV-1 infection in LAPTM5-expressing CD4⁺ T cells.

a–c TZM-bl cells were transfected with FLAG-tagged LAPTM5 or mock expression constructs to express LAPTM5 at similar levels as in MDMs. The same number of TZM-bl cells or MDMs was lysed for western blotting to assess LAPTM5 and GAPDH expression (a). LAPTM5-positive or -negative-TZM-bl cells were infected with 100 ng of wild-type or Vpr-defective HIV-1 (b, c) for 3 days. At 1 dpi, cells were treated with raltegravir (2 μM) to block subsequent rounds of infection. HIV-1 capsids or Env-containing virions in culture were measured by p24 or gp120 ELISA, respectively. Data are plotted as mean ± SEM of three independent experiments. Cells were lysed for western blotting to assess gp120, Gag, LAPTM5 and GAPDH expression. d, e CD4⁺ T cells were stimulated and transduced with tetracycline-controlled LAPTM5 or mock expression lentiviral vectors to produce LAPTM5 at similar levels as in MDMs. The same number of CD4⁺ T cells or MDMs was lysed for western blotting to assess LAPTM5 and GAPDH expression (d). LAPTM5-positive or -negative-CD4⁺ T cells were infected with 100 ng of dual-trophic wild-type or Vpr-defective HIV-1_89.6 for 6 days (e). Viral production was measured by p24 ELISA at the indicated time points. f Working model of how Vpr counteracts LAPTM5 to promote HIV-1 infection. Data are plotted as mean ± SEM of three independent experiments. All western blot data are representative of three independent experiments; their full-size images are presented in the Source Data.