Fig. 6: Vpr counteracts the LAPTM5-mediated degradation of HIV-1 Env.
From: Vpr counteracts the restriction of LAPTM5 to promote HIV-1 infection in macrophages
a–c Lentiviral shRNA-transduced MDMs were infected with 10 or 100 ng of wild-type or Vpr-defective HIV-1 for 6 days. Before infection, the aliquoted shRNA-transduced MDMs were lysed for western blotting (a) to assess LAPTM5 and GAPDH expression. At 2 dpi, cells were treated with raltegravir (2 μM) to block subsequent rounds of infection. HIV-1 capsids or Env gp120-containing virions (b, c) in culture were measured by p24 or gp120 ELISA, respectively. Data are plotted as mean ± SEM of three independent experiments. d Lentiviral shRNA-transduced MDMs were infected with 500 ng of wild-type or Vpr-defective HIV-1AD8 in the presence or absence of a lysosomal inhibitor (at 50 nM treatment for the last 24 h). At 2 dpi, cells were treated with raltegravir (2 μM) to block subsequent rounds of infection. Eight days after infection, cells and isolated lysosomes were lysed for western blotting to assess LAMP1, LAPTM5, Gag, gp120, and GAPDH expression. All western blot data are representative of three independent experiments; their full-size images are presented in the Source Data.
