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. 2023 Mar 28;14(2):e00166-23. doi: 10.1128/mbio.00166-23

FIG 2.

FIG 2

SERINC5 virion association renders the particles defective in THP-1. (A) Infectivity of HIV-1 SERINC5+/− virions in THP-1-derived target macrophages that were primed with either conditioned media (CM) or VSV-G-enveloped vesicles prior to infection with Nef-defective HIV-1. Infectivity was normalized to RT units. Vesicles were produced by transfection of envelope glycoprotein along with empty vector (SERINC5−) or vector expressing SERINC5 (SERINC5+). The values are means (n = 3) and SD. An unpaired t test was used. (B) THP-1-derived macrophages infected with HIV-1 produced from HEK293T by cotransfecting pMD2.G, NL4-3, Luciferase Env(−) R(−), SERINC5, or equivalent empty vector, along with HIV-1 Nef-HA/MLV-HA-GlycoGag/empty vector. Infectivity was normalized to RT units. Lower panel, Western blot showing C-terminally FLAG-tagged SERINC5 and HA-tagged Nef Lai and GlycoGag with the corresponding β-actin from virus-producing cell lysate (HEK293T). The values are means (n = 3) and SD. An unpaired t test was used. (C) THP-1-derived macrophages were infected with HIV-1 produced as in panel B with HIV-1 Lai Nef or indicated Nef mutants. Infectivity was normalized to RT units. Lower panel, Western blot showing C-terminally FLAG-tagged SERINC5 and HA-tagged WT and mutants of Nef Lai with the corresponding β-actin from cell lysate (HEK293T). The values are means (n = 3) and SD. An unpaired t test was used. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; HA, hemagglutinin; MLV, murine leukemia virus; ns, not significant.