Fig. 1.

IAV NS2 protein interacts with CHD3. a NS2 binds CHD3 in M2H assay. COS-1 cells were cotransfected with the plasmids pACT–cCHD3, pBIND–NS2, and pG5luc, and then cell lysates were subjected to luciferase activity assays via M2H 24 h later. pBIND-Id and pACT-MyoD were used as positive controls, and pACT and pBIND as negative controls. The results are presented as the mean ± SD (**p < 0.01, n = 3). b NS2 interacts with CHD3 in Co-IP assay. Myc-NS2 and Flag-NLS-cCHD3 were cotransfected into COS-1 cells. 36 h after transfection, the cells were lysed, and immunoprecipitation was performed using an anti-Flag monoclonal antibody. The immunoprecipitated proteins were assayed with an anti-Myc polyclonal antibody. c NS2 interacts with CHD3 in GST pull-down assay. GST-tagged NS2 was expressed in bacteria. The cell lysates containing Flag-NLS-cCHD3 were mixed with beads to which GST-NS2 or GST alone was bound. The samples were analyzed using an anti-Flag monoclonal antibody. d NLS–cCHD3 interacted with NS2 in the nucleus by IFA assay. COS-1 cells on coverslips were transfected with plasmids coding the NLS–cCHD3 and NS2, and analyzed by IFA using antibodies against Myc (rabbit) and Flag (mouse). Colocalization was analyzed using Image J and the coefficients confirmed the strong colocalization