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. 2015 Mar 10;290(17):10919–10933. doi: 10.1074/jbc.M114.630095

FIGURE 3.

FIGURE 3.

Staining for surface Env and BST2 in the presence of Vpu-mutants also reveals the displacement effect. A, HeLa P4-R5 cells were transfected with the indicated proviral Vpu-mutant plasmids and the next day stained for BST2 (in red) and Env (in green), then fixed and imaged. The merged image is shown on the right with BST2-Env overlap in yellow. Wild-type (WT) Vpu and the S52N,S56N mutant are each able to displace BST2 from Env (compared with ΔVpu), and this activity is lost for W76G and for the Vpu transmembrane domain mutant A10F,A14F,A18F, which is unable to interact with BST2. B, Pearson's correlation coefficients for the colocalization of BST2 and Env along the plasma membrane in the presence of Vpu or mutants from the experiment shown in A. At least 20 cells were assessed for each condition, using image planes 1–3 μm above the cover glass. Error bars are the standard deviations. p values for unpaired two-tailed t tests were as noted on the graph and for ΔVpu versus WT and S52N,S56N were <0.0001; for ΔVpu versus W76G, p = 0.0810; for WT versus W76G, S52N,S56N, S52N,S56N + W76G, and A10F,A14F,A18F, p < 0.0001; for W76G versus S52N,S56N, p < 0.0001; for W76G versus S52N,S56N + W76G, p = 0.0037; for W76G versus A10F,A14F,A18F, p = 0.0554; for S52N,S56N versus A10F,A14F,A18F, p < 0.0001; and for S52N,S56N + W76G versus A10F,A14F,A18F, p = 0.4075.