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. 2011 Feb 10;8:9. doi: 10.1186/1742-4690-8-9

Figure 2.

Figure 2

Expression of an F-box-deficient β-TrCP impairs Vpu antagonism and surface downregulation of CD4 and CD317, but not depletion of CD317. 293TCD4 cells were transfected to express HIV-1Δnef Δvpu GFP, HA-CD317, and Vpu together with either Myc-tagged β-TrCP1 wt or β-TrCP1ΔF. Two days post-transfection, supernatants and cells were analyzed as described in the legend to Figure 1. (A) The yield of infectious HIV-1 and (B) cell-associated levels of HA-CD317, Vpu, β-TrCP-Myc, and MAPK were determined. Western blots shown represent samples run on the identical gel with gaps indicating areas where non-informative lanes were omitted. (C, D) Relative surface levels of CD317 and CD4 on HIV-1-expressing (GFP-positive) cells. Shown are arithmetic means ± SEM of 3 to 6 independent experiments. Student's t-test: * p < 0.05, *** p < 0.001. (E, F) Cells were processed for microscopic analysis to visualize and quantify the frequency of cells with p24CA+ aggregates in relation to co-expressed β-TrCP1 wt or β-TrCP1ΔF. Values are from one experiment and representative for two analyses. Arrow heads indicate plasma membrane and intracellular p24CA+ aggregates. Scale bar: 10 μm.