Fig 3.

Nucleozin treatment does not disrupt RNPs. 293T cells were transfected with plasmids expressing GFP alone or GFP-NP, incubated for 24 h, and infected with the WT virus or mock infected. At 6 hpi, one set of cells was treated with 1 μM nucleozin (NCZ). Cell lysates prepared at 8 hpi were analyzed by Western blotting for the indicated polypeptides after GFP-Trap affinity selection into supernatant and bound fractions. Sample loading is such that the supernatants are equivalent to 1/10 of the bound fractions. RNA was analyzed by primer extension for segment 3 vRNA and 5S rRNA before or after GFP-Trap selection. The experimental procedure was performed twice. Cont., control.