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. 2007 Jan 24;81(7):3428–3436. doi: 10.1128/JVI.02303-06

FIG. 2.

FIG. 2.

Effects of NS1 and NS2 on STAT1 and STAT2 in 293T cells. (A) Ev, or NS1 or NS2 cDNA, was transfected into 293T cells. Cells were treated with proteasome inhibitors (MG132 and LLnL) 4 h prior to lysis to confirm NS1 expression (left panel) or lysed in 8 M urea to analyze NS2 expression (right panel). (B and C) Cells were transfected with cDNA encoding FLAG-STAT1 (B) or STAT2 (C) in the presence of NS1, NS2, or both NS1 and NS2. Membranes were probed with anti-FLAG to detect STAT1 or anti-STAT2 (upper panels) and reprobed with anti-γ-tubulin to confirm equal protein loading in all lanes (lower panels). (D) Loss of STAT2 protein by NS1 and NS2 can be prevented by blocking proteasomal activity. 293T cells were transfected with vectors expressing STAT2 in the absence and presence of NS1/2. Cells were treated with and without proteasome inhibitors (MG132 and LLnL) 3 h before lysing. STAT2 levels were analyzed by immunoblotting membranes with anti-STAT2 (upper panels). Membranes were reprobed with anti-γ-tubulin (lower panels).