Figure 2.

Investigation of the interaction domain of NS2 and the interaction between NS2 and endogenous H1C. (A) Human embryonic kidney 293 T cells were transfected with Flag-H1C and RFP-NS2 or its mutants, and a co-immunoprecipitation (Co-IP) was performed to detect interactions using the anti-flag antibody. An anti-RFP antibody was used to detect pDsRed-NS2 or its mutants. (B) HeLa cells cultured on slides were co-transfected with pEGFP-H1C and pDsRed-NS2, pDsRed-NS2-NT, or pDsRed-NS2-CT, and confocal microscopy was performed 24 h later to detect the co-localization. (C) A549 cells were infected with influenza virus for 10 h (WSN,MOI = 10), and an IP experiment was performed using an anti-NS2 antibody to detected the interaction between NS2 and endogenous H1C. Histone H4 (H4) served as a negative control. (D) A549 cells were treated as described above, and confocal microscopy was performed using the anti-NS2 mouse polyclonal antibody and anti-H1C rabbit polyclonal antibody followed by immunostaining with FITC-labeled goat anti-mouse secondary antibody and Cy3-labeled goat anti-rabbit antibody to detect its co-localization.