Fig. 6

Impaired BST2-binding Vpu TMD mutants display defects in cellular distribution. a Enhancement of virus release by Vpu TMD and E28A/L33A mutants. A representative Western blot showing the amount of virion-associated p24 released into the supernatant (virion) and Gag products (p24 and p55) in the cell lysate. Below the blot are values indicating the efficiency of virus release (quantified and expressed as mentioned in Fig. 4 legend). b–d Vpu TMD mutants are defective for TGN localization. b HeLa cells transfected with either WT, E28AL/L33A, A18L or A10L/A18L Vpu mutants were co-stained with anti-TGN46 (red, for TGN) and anti-Vpu (green) Abs as well as with DAPI (grey, for nucleus). Shown are representative confocal microscopy pictures for each of the Vpu mutants. c, d Quantification of the co-staining of anti-Vpu and anti-TGN46 Abs obtained from at least 50 distinct transfected cells per mutant. Shown are Pearson correlation coefficients (PCC) for each mutant (c) as well as Vpu distribution beyond the TGN (d). The percentage of Vpu distributing beyond the TGN was determined by calculating the ratio of the intensity of Vpu not co-staining with TGN versus total Vpu intensity in transfected cells. The white bars in B represent a distance of 10 µm and the horizontal lines (c, d) represent mean values of the PCC (c) and percentage of Vpu distributing beyond the TGN (d). Statistical analyses were performed using Mann–Whitney test