Fig. 4
Maintenance of HIV-1-like Vpu function in GU1N infected AGMs. a Alignment of Vpu sequences derived from GU1N-infected AGMs at 121 (2012) and 223 wpi (2014) and primary or synthetic (Sel1 and Sel2) Vpus chosen for functional analysis. Orange dots highlight mutations found in all three AGMs. Dashes indicate gaps introduced to optimize the alignment. Numbers specify the animal, year of plasma isolation, and number of the respective Vpu sequences. Black indicates changes in all, blue in ≥50% and gray in at least two sequences. b Expression of AU1-tagged SIVgsn Vpu proteins. HEK293T cells were transfected with expression plasmids encoding the indicated AU1-tagged Vpus and eGFP. Mock transfected cells were used as negative controls. GAPDH expression levels were analyzed to control for loading. c Infectious virus (left) and p24 antigen (right) yield from HEK293T cells cotransfected with an HIV-1 NL4-3 Δvpu construct and vectors expressing the indicated vpu alleles in combination with increasing amounts of plasmids expressing AGM tetherin. Shown are average values derived from three experiments relative to those obtained in the absence of tetherin (100%). d Vpu-dependent reduction of CD4, NTB-A, CD1d, and AGM tetherin surface expression in HEK293T cells relative to those measured in cells transfected with the eGFP only control vector. Values in panels d and e represent mean (+SEM) derived from three independent experiments. e AGM-derived Vpu proteins suppress NF-κB activity. The effects were measured as described in the legend to Fig. 1c
