Nedd4 binding to the PPxY prevents cytoplasmic translocation of Daxx by protein VI.

<p>Endogenous Daxx in U2OS cells was detected following Mock transfection (A), transfection with VI-wt (B) or after cotransfection of VI-wt with Nedd 4.2 (C) or catalytical inactive Nedd 4.2-TD (D), transfection of VI-M1 (E) or VI-M1 cotransfected with Nedd 4.2. (F) as indicated to the left of each row. Daxx was stained with Ab against the endogenous protein (first column), VI was detected using the RFP signal (second column) or GFP for Nedd4 (third column). An overlay is shown in the last column showing Daxx in blue, protein VI in red, Nedd4 in green and the nucleus in grey. Phenotypes shown in representative cells were observed in all transfected cells. Colocalization of VI, Nedd4.2 and PML is shown in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1002549#ppat.1002549.s005" target="_blank">Figure S5</a>.