Skip to main content
. 2019 Aug 28;93(18):e00322-19. doi: 10.1128/JVI.00322-19

FIG 4.

FIG 4

NDV V interacts with MAVS. (A) HEK-293T cells were cotransfected with Flag-V and either empty vector or HA-MAVS. At 24 hpt, cells were harvested, immunoprecipitated with anti-HA antibody, and further detected using immunoblot analysis with anti-HA or anti-Flag antibody. Expression levels of the proteins were analyzed by immunoblot analysis of the lysates with anti-HA, anti-Flag, or anti-β-actin antibody. (B) HEK-293T cells were cotransfected with HA-MAVS and either empty vector or Flag-V. At 24 hpt, cells were harvested, immunoprecipitated with anti-HA antibody, and further detected using immunoblot analysis with anti-HA or anti-Flag antibody. Expression levels of the proteins were analyzed by immunoblot analysis of the lysates with anti-HA, anti-Flag, or anti-β-actin antibody. (C) HEK-293T cells were cotransfected with HA-MAVS and either empty vector or Flag-V, -P, or -W. At 24 hpt, cells were harvested, immunoprecipitated with anti-Flag antibody, and further detected using immunoblot analysis with anti-HA or anti-Flag antibody. Expression levels of the proteins were analyzed by immunoblot analysis of the lysates with anti-HA, anti-Flag, or anti-β-actin antibody. (D) HeLa cells were mock treated or infected with NDV at an MOI of 1. At 24 hpt, cells were harvested, immunoprecipitated with anti-IgG or MAVS antibody, and further detected using immunoblot analysis with anti-V or anti-MAVS antibody. Expression levels of the proteins were analyzed by immunoblot analysis of the lysates with anti-V, anti-MAVS, or anti-β-actin antibody. (E) HeLa cells were mock treated or infected with NDV at an MOI of 1. Cells were harvested at 6, 12, 18, and 24 hpi and detected using IF assay with anti-MAVS or anti-V antibody. Shown is an intensity profile of the linear region of interest (ROI) across the HeLa cell costained with V and MAVS. (F) Schematics of a series of Flag-tagged truncated V constructs. (G) HEK-293T cells were cotransfected with HA-MAVS and either empty vector, WT, or truncated Flag-V (ΔDM1, ΔDM2, and ΔDM3). At 24 hpt, cells were harvested, immunoprecipitated with anti-IgG or anti-Flag antibody, and further detected using immunoblot analysis with anti-HA or anti-Flag antibody. Expression levels of the proteins were analyzed by immunoblot analysis of the lysates with anti-HA, anti-Flag, or anti-β-actin antibody. (H) Schematics of a series of HA-tagged truncated MAVS constructs. (I and J) HEK-293T cells were cotransfected with Flag-V and either empty vector or truncated HA-MAVS aa 1 to 180, aa 180 to 360, and aa 360 to 540 (I) or aa 1 to 360, aa 180 to 540, and ΔTM (J). At 24 hpt, cells were harvested, immunoprecipitated with anti-Flag or anti-HA antibody, and further detected using immunoblot analysis with anti-HA or anti-Flag antibody. Expression levels of the proteins were analyzed by immunoblot analysis of the lysates with anti-HA, anti-Flag, or anti-β-actin antibody.