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. 2011 Aug;85(16):8056–8068. doi: 10.1128/JVI.01878-10

Fig. 5.

Fig. 5.

Recombinant NV NS6/7 protein and FCV NS6/7 protein coprecipitate with nucleolin. (A) Recombinant Norwalk virus protein (NV rNS6/7) coupled to Ni-NTA resin or Ni-NTA resin alone was incubated with HeLa or CrFK cell extracts as indicated. After several washes, the precipitated proteins were analyzed by SDS-PAGE, transferred to a nitrocellulose membrane, and analyzed by Western blotting using human antinucleolin (right) or anti-NV rNS6/7 (left) antibody. Molecular masses of nucleolin and NV NS6/7 are indicated to the right of the panels. PD, pulldown. (B) Antinucleolin or antiactin antibody was incubated with total extracts from FCV-infected cells, and immunocomplexes were immobilized in protein G-agarose resin, precipitated, and analyzed by SDS-PAGE. The transferred proteins were analyzed by Western blotting using human antinucleolin (left) or anti-FCV NS6 (right) antibody. Co-IPP, coimmunoprecipitation. Arrows indicate migration of nucleolin and NV rNS6/7. Input cell extracts are shown in the third lane of each panel.