Fig. 2.

DENV replication and translation are not affected by inhibitors of cap-dependent translation, (A) DENV is replicated in the presence of rapamycin, LY294002 or wortmannin. Cells were exposed to DENV for 1 h in the presence of inhibitors, washed, and incubated with rapamycin (250 ng/ml), LY294002 (40 μM) or wortmannin (1 μM) for 24 h. Supernatants were collected and titered by plaque assay (PFU/ml), (B) DENV RNA is translated in the presence of LY294002 or wortmannin. Cells were prepared as described above. At 12 h post-infection, cells were metabolically labeled and analyzed by SDS-PAGE. ImageQuant software (Molecular Dynamics) was used to quantify the DENV protein NS5 (arrow) and representative cellular proteins.