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. 2010 Feb 10;84(8):4089–4094. doi: 10.1128/JVI.01549-09

FIG. 4.

FIG. 4.

Susceptibilities of WT and mutant CD317 proteins to Vpu-mediated downregulation from the cell surface upon HIV-1 infection. 293T cells stably expressing pcDNA3.1 (vector), WT HA-CD317, or HA-CD317 mutants were infected with VSV G-pseudotyped WT HIV-1 or HIV-1 Δvpu. Seventy-two hours postinfection, cells were analyzed by flow cytometry using surface CD317 staining (anti-CD317 monoclonal antibody from Chugai Pharmaceuticals) to quantify HA-CD317 surface expression levels and concurrent intracellular p24CA staining to identify productively infected cells. Uninfected cells and HA-CD317-expressing cells infected in the presence of the reverse transcriptase inhibitor efavirenz (1 μM) served as negative controls (not shown). Values depicted are the arithmetic means of triplicates ± standard errors of the means from three independent experiments. (A) The relative cell surface expression levels of HA-CD317 and mutants are depicted, with values for the HIV-1 Δvpu-infected cells set to 100%. (B) Quantification of infectious HIV-1 in the cell supernatant at the time of cell harvest for flow cytometry.