(
A) Conservation of PPP2R5A subunit degradation by HIV-1 Vif variants. 293T cells stably expressing HA-PPP2R5A were transfected with the indicated Vif variants and analysed by intracellular flow cytometry for HA 36 hr post-transfection. The median fluorescence intensity of the transfected population is shown as a proportion of median fluorescence intensity of the untransfected population for each condition, normalized to the empty vector control. Mean values and standard errors are shown. (
B) Conservation of PPP2R5A-E subunit degradation by phylogenetically diverse lentiviral Vif variants. 293T cells stably expressing different HA-tagged PPP2R5A-E subunits were transfected with the indicated Vif variants and analysed by intracellular flow cytometry for HA 36 hr post-transfection. Median fluorescence intensity of the transfected population is shown as a proportion of median fluorescence intensity of the untransfected population for each condition, normalized to the empty vector control. Each datapoint represents a different Vif variant. (
C) Representative data for primate lentiviral Vif degradation of PPP2R5B. Data shown in
Figure 7B was acquired in several experiments, one example is shown here. Histograms show GFP positive (transfected, red shading) and negative (untransfected, blue line) cells. MFI values are shown for GFP positive (red) and negative (blue) cells.