FIG. 2.

The NES21 and NES22 substitutions within the NS2 NES site prevent the interaction of NS2 with Crm1 in vivo. 293T cells were transfected with pdBMVp, pdBMV-NES21, pdBMV-NES22, or no DNA (mock) and further incubated for 48 h. (A) Whole-cell lysates were first immunoprecipitated with an NS2-specific antiserum and then immunoblotted with anti-Crm1 antibodies. As controls, nonimmunoprecipitated lysates (10% of the amount used for panel A) were analyzed by immunoblotting with either the anti-Crm1 (B) or anti-NS2 (C) antibodies. Proteins were separated on SDS-containing bipartite 8 and 12% polyacrylamide gels. Immunoblots were visualized by chemiluminescence.