Figure 7

The nuclear export -deficient mutants of the full length influenza A/Udorn/72 NS1 protein bind in vitro -translated CPFS30 protein in a GST pull down experiment. [³⁵S]-labeled and in vitro-translated CPFS30 was allowed to bind to E. coli-expressed and Sepharose-immobilized GST, GST-NS1 A/Udorn/72 wt, GST-NS1 F138A, L141A, GST-NS1 L144A, L146A and GST-NS1 A149V at +4°C for 1 h. Sepharose-bound proteins were dissolved in Laemmli sample buffer, separated on 12% SDS-PAGE and autoradiographed. Input, lane C, was 1:20 of the amount of [³⁵S]-labeled protein that was used in binding experiment. M indicates the Sepharose matrix-bound proteins. The same samples were also separated on 12% SDS-PAGE and stained with Coomassie Blue to visualize the amount of Sepharose-immobilized GST and GST-NS1 fusion proteins.