Fig. 2: LAPTM5 restricts HIV-1 infectivity.

a HeLa cells were transfected with FLAG-tagged LAPTM5 or mock expression constructs. At 24 h after transfection, cells were infected with 100 ng of Vpr-defective HIV-1_{NL4-3.Luc.R-E-} (VSV-G). At 2 dpi, cells were lysed to measure luciferase reporter activity, and western blotting was performed to assess exogenous protein expression. Data are plotted as mean ± SEM of three independent experiments. b HeLa cells were cotransfected with FLAG-tagged LAPTM5 or mock expression constructs along with a Vpr-defective pNL4-3.GFP.R⁻E⁻ reporter vector combined with various expression constructs of HIV-1 ×4-, R5-trophic, or dual-trophic Env or VSV-G as indicated. Two days after transfection, TZM-bl indicator cells were used to measure HIV-1 infectivity. *P < 0.05, **P < 0.01; NS, not significant (two-tailed, unpaired Student’s t-test), data are plotted as mean ± SEM of three independent experiments. c, d HeLa cells were cotransfected with non-tagged LAPTM5 or mock expression constructs along with Vpr-defective HIV-1 proviral vectors (NL4-3 Vpr- or BaL Vpr-) and a monitoring expression plasmid of GFP in the presence or absence of a FLAG-tagged Vpr expression vector derived from NL4-3. Two days after transfection, TZM-bl indicator cells were used to measure HIV-1 infectivity (c). Cells were lysed for western blotting to assess LAPTM5, Vpr, GFP, and GAPDH expression (d). 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.