Fig 2.

Vpu is necessary and sufficient for disruption of IRF3-dependent signaling and IRF3 depletion. (A) Wild-type and Vpu-deficient proviral mutants of JR-CSF, pNL4-3, AD8, and YU2 were transfected and tested for signaling in response to SeV as described for Fig. 1A; constructs were tested for expression of Vpu, Vpr, Vif, HIV-1 Gag, or actin by immunoblotting. *, P < 0.05; **, P < 0.01. (B) Cells were transfected as described for Fig. 1A and immunoblotted for IRF3, Vpu, and actin. (C) Cells were treated as described for panel A, with Vpu added in trans in increasing doses along with the Vpu-deficient pNL4-3 proviral construct. Luciferase reporter gene experiments were repeated 3 or more times, and representative immunoblot analyses are shown.