A36 phosphorylation and recruitment of Nck displaces septins. (A) Representative images showing the association of SEPT7 with CEV in WR-infected HeLa cells in the presence or absence (CTRL) of PP1. The graph shows the percentage of CEV in WR- and YdF-infected cells recruiting SEPT7. (B) The percentage of CEV recruiting SEPT7 in HeLa cells infected with vaccinia expressing A36 (WR), A36-Y132F (Y132F), A36-Y112F (Y112F), and A36-YdF (YdF). (C) Immunofluorescence images showing the recruitment of SEPT7 to CEV in Nck parental and knockout MEFs infected with WR and YdF virus for 20 h. The graph shows the percentage of CEV with SEPT7 for the indicated viruses in the presence (+/+) or absence (−/−) of Nck. (D) Quantification of the percentage of CEV recruiting SEPT7 in Nck knockout MEFs stably expressing GFP-Nck or the indicated function disrupting SH3 domain point mutants. Error bars represent SEM from three independent experiments in which a total of 900 CEVs were analyzed in 30 cells. Bars, 5 µm. ***, P < 0.001; ****, P < 0.0001.