FIG. 8.

Effect of poliovirus 2A protease on the relocation of 3×GFP-NLS from the nuclei of uninfected cells. In the upper set of panels (a to c), HeLa-3E cells were untreated (a) or transfected with plasmids expressing either the wild-type poliovirus 2A (b) or 2A harboring a protease inactivating H20N mutation (c). The pictures were taken 12 h after the transfection. In the lower set of panels (d to k), twofold-diluted S15 extracts from HeLa cells expressing wild-type (d to f and h to j) and mutant (g and k) 2A were applied to the permeabilized uninfected HeLa-3E cells for 1 h at 37°C. When used, zVAD.fmk and MPCMK were at 5 and 50 μM, respectively, and preincubated with the extracts for 20 min at 4°C.