Fig. 3: LAPTM5 restricts HIV-1 infectivity and Vpr counteracts the restriction.
From: Vpr counteracts the restriction of LAPTM5 to promote HIV-1 infection in macrophages
a HeLa cells were cotransfected with FLAG-tagged LAPTM5 or mock expression constructs at the indicated doses along with wild-type or Vpr-defective HIV-1AD8 proviral vectors. Two days after transfection, TZM-bl indicator cells were used to measure HIV-1 infectivity. Data are plotted as mean ± SEM of three independent experiments. b HeLa cells were cotransfected with a bicistronic construct containing an IRES to coexpress LAPTM5 and GFP (LAPTM5-IRES-GFP) with or without FLAG-tagged Vpr expression vectors derived from different HIV-1 isolates. Two days after transfection, cells were lysed to assess LAPTM5, Vpr, GFP, and GAPDH expression by western blotting. c HeLa cells were cotransfected with GFP-tagged LAPTM5 or mock (GFP) expression constructs with or without a FLAG-tagged Vpr expression vector in the presence or absence of MG132 (1.5 μM treatment for the last 12 h). Two days after transfection, cells were lysed to assess LAPTM5-GFP, GFP, Vpr, and GAPDH expression by western blotting. d HeLa cells were cotransfected with GFP-tagged LAPTM5 or mock (GFP) expression constructs with or without a FLAG-tagged Vpr expression vector in the presence of siRNA against DCAF1 or control siRNA. Two days after transfection, cells were lysed to assess LAPTM5-GFP, GFP, DCAF1, Vpr, and GAPDH expression by western blotting. e Lentiviral shRNA-transduced MDMs were infected with 500 ng wild-type or Vpr-defective HIV-1AD8 in the presence or absence of EFV (0.3 μM). At 6 dpi, cells were lysed to assess LAPTM5, DCAF1, Gag, and GAPDH expression by western blotting. All western blot data are representative of three independent experiments; their full-size images are presented in the Source Data.
