Cycloheximide pretreatment blocks rNef-induced STAT1 activation.

Western blot analysis of phosphotyrosine-STAT1, STAT1, IRF-1, or β-tubulin on total cell extracts of MDMs pretreated or not with 5 μg/mL cycloheximide and incubated for 2 hours with either 100 ng/mL rNef or 100 IU/mL human IFN-γ for 2 hours. As controls, the analyses were carried out on cell extracts of MDMs treated either for 2 hours with 100 ng/mL rNef or with cycloheximide only. Cell extracts were obtained by pooling simultaneous cell cultures from 4 healthy donors. Specific signals are indicated on the left side; molecular size markers (in kilodaltons) are reported on the right.