Journal: Virus Research
Article Title: Foot-and-mouth disease virus 3D polymerase antagonizes the interferon signaling pathway by blocking STAT2 nuclear translocation
doi: 10.1016/j.virusres.2025.199671
Figure Lengend Snippet: FMDV 3D protein interacts with STAT2. (A) HEK-293T cells were co-transfected with Flag-3D and various Myc-tagged innate immune molecule-expressing plasmids (JAK1, TYK2, STAT1, STAT2, or IRF9). At 36 hpt, the cell lysates were subjected to Co-IP assay analysis. Immunoprecipitated proteins and whole-cell lysates (WCL) were analyzed by western blotting using specified antibodies. ( B) HEK-293T cells were co-transfected with Myc-STAT2 and either an empty vector or Flag-3D expressing plasmids for 36 h. Cell lysates were immunoprecipitated with anti-Myc or control IgG antibodies and analyzed by western blotting. ( C) HEK-293T cells were co-transfected with Flag-3D along with Vec or Myc-STAT2. At 36 hpt, cell lysates were subjected to Co-IP assay. Immunoprecipitated proteins and WCL were analyzed by western blotting. (D) PK-15 cells were transfected with Flag-3D or empty vector plasmids. At 36 hpt, cell lysates were immunoprecipitated with anti-Flag antibodies and analyzed by western blotting. (E) PK-15 cells were mock-infected or infected with FMDV for 12 h, cell lysates were immunoprecipitated with anti-3D antibodies and analyzed by western blotting with the indicated antibodies. (F) PK-15 cells were transfected with porcine HA-STAT2 expressing plasmids for 24 h, then mock-infected or infected with FMDV (MOI = 0.1) for 10 h. Colocalization of HA-STAT2 (red) and FMDV 3D (green) was assessed by immunofluorescence assay (IFA). Nuclei were counterstained with DAPI (blue).
Article Snippet: The commercial antibodies used in this study include: anti-Flag mouse Ab (Sigma, F1804), anti-Myc mouse Ab (Sigma, M5546), anti-HA mouse Ab (Sigma, H9658), and anti-GAPDH mouse Ab (Abclonal, AC002), anti-JAK1 rabbit Ab (Cell Signaling Technology, 3332), anti-TYK2 rabbit Ab (Cell Signaling Technology, 9312), anti-STAT1 rabbit Ab (Cell Signaling Technology, 9172), anti-STAT2 rabbit Ab (Cell Signaling Technology, 4594), anti-p-STAT2 rabbit Ab (Cell Signaling Technology, 4441), anti-IRF9 rabbit Ab (Cell Signaling Technology, 76,684).
Techniques: Transfection, Expressing, Co-Immunoprecipitation Assay, Immunoprecipitation, Western Blot, Plasmid Preparation, Control, Infection, Immunofluorescence
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