FIG 7.

Vimentin dynamics affect the synthesis of nonstructural proteins rather than their degradation. (A) Western blot of A549 cells infected with CVB3 for 3, 4, 5, or 6 h with or without IDPN treatment. Virus (4.43 × 10⁸ PFU/ml) was bound on ice for 1 h. After washing the excess virus away, infection was allowed to proceed for the indicated time. IDPN was added after ice binding and left until the end of the experiment. eIF4G, 3D, 2A, and VP1 were visualized using antibodies against the proteins. Shown is a representative image of two replicates. (B) Western blot showing the effect of 1.5% IDPN, 7 μM bortezomib, 200 μM calpain inhibitor 1, 100 μg/ml puromycin, 50 μM VER-155008, or 0.1 μM geldanamycin on CVB3 infection. Virus (4.43 × 10⁸ PFU/ml) was bound on ice for 1 h. After washing the excess virus away, infection was allowed to proceed for 5.5 h. Other drugs were added after ice binding except calpain inhibitor, bortezomib, and puromycin, which were added at 2 h p.i. All drugs were left until the end of the experiment. Visualization of VP1 and P1 is on the left. Merged image of 2A and VP1 labeling is on the right. Results are representative of at least two separate experiments.