Figure 4
From: ASFV pD345L protein negatively regulates NF-κB signalling by inhibiting IKK kinase activity
pD345L interacts with IKKα and IKKβ. A coimmunoprecipitation assay was performed with whole cell lysates prepared with 293 T cells co-transfected with Flag-D345L and HA-IKKα for 36 h with Flag antibody (A), HA antibody (B) or control IgG. The immunocomplexes were analysed by immunoblotting with the indicated antibodies. C and D The experiment was performed as for Panel A or B except using HA-IKKβ. E A coimmunoprecipitation assay was performed with whole-cell lysates prepared with PAM cells infected with ASFV (MOI 1) for 24 h with pD345L antibody. The immunocomplexes were analysed by immunoblotting with the indicated antibodies. F and G Diagrams of truncated IKKα and IKKβ. Both IKKα and IKKβ contain an N-terminal kinase domain (KD), a C-terminal helix–loop–helix domain (HLH) and a leucine zipper domain (LZ), as indicated. The experiment was performed as for A, except that HA-D345L and Flag-tagged IKKα, IKKα (1-371), IKKα (371-500), IKKα (500–745), empty vector (H), or Flag-tagged IKKβ, IKKβ (1–399), IKKβ (399–577), IKKβ (577–756), empty vector (I) were transfected as indicated.