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. 2009 Jun 12;5(6):e1000473. doi: 10.1371/journal.ppat.1000473

Figure 2. The dsRNA-binding activity of the NS1 protein mediates interaction with PKR.

Figure 2

(A) Human A549 lung epithelial cells were mock treated or infected with WT virus or the NS1 mutant viruses #1, #2, #3, #4, #6 or #7 at an MOI of 1. Lysates were subjected to immunoprecipitation (IP) with NS1 antiserum (α) or pre-serum (ctrl). The precipitated complexes were analyzed by immunoblotting with antibodies specific for PKR (upper panel) and NS1 (lower panel). Whole cell lysates were also analyzed with antibodies specific for PKR, tubulin and NS1 as indicated. (B) A549 cells were mock treated or infected with WT virus at an MOI of 1. Lysates were prepared that were either mock treated (−) or treated with the indicated amounts of RNase III for 10 minutes at 33°C prior to immunoprecipitation with NS1 antiserum (α) or pre-serum (ctrl). The precipitates were analyzed as described in panel 2A (panel to the left). Aliquots of the lysates were also analyzed directly by immunoblotting for PKR, tubulin and NS1 as indicated (panels to the right).