Fig 2.

Actin-based motility of ECTV is via an N-Wasp-dependent pathway. (A) BSC-1 cells were infected with ECTV or VACV at an MOI of 1 and fixed at the indicated time points. Cells were stained with phalloidin and assessed for the presence of actin comets (n = 50 per time point, in duplicate). The percentage of cells with actin tails was calculated. Error bars indicate standard errors. (B) HeLa cells were infected with ECTV or VACV, fixed, and stained for immunofluorescence assays with anti-B5 (NPC) (red) or phalloidin (green). Arrowheads indicate a WV associated with an actin comet. ECTV-infected cells were also probed with anti-B5 (NPC) (red), anti-A36-Y112 (green), and DAPI (blue) (C) or with phalloidin (red), an antiphosphotyrosine antibody (4G10; green), and DAPI (blue) (D). (E) N-Wasp^−/− MEFs were mock transfected or transfected with an N-Wasp-GFP expression vector (green). At 24 h posttransfection, cells were infected with ECTV and fixed. For immunofluorescence assays, fixed cells were stained with phalloidin (red) and DAPI (blue). Arrowheads indicate WV (C) or WV associated with an actin comet (D and E). Bars, 10 μm.