In vitro and in vivo interaction between NS1 and Gas8. A GST pull-down analysis. Lysates from cells expressing myc-Gas8 were mixed with GST or GST-NS1, and bound proteins were analyzed by anti-myc western blot. Left lane, cell lysate; middle and right lanes, proteins bound to GST-NS1 and GST, respectively. Gas8 was precipitated by a GST-NS1 fusion protein but not by GST. (B) Western blot of NS1 and Gas8 co-IPs. Left: Cells were co-transfected with a plasmid expressing myc-tagged NS1 and either a plasmid expressing FLAG-tagged Gas8 or the empty vector. Cell lysates were IPed with anti-FLAG antibody (left two lanes) or run on a gel without immunoprecipitation (right two lanes). Samples were probed with anti-myc and anti-FLAG antibody. Right: IP with anti-myc antibody and IB with anti-FLAG and anti-myc antibody. The results showed that NS1 could be precipitated with Gas8 and Gas8 could also be precipitated with NS1.