Figure 6
Virus triggers PC cytoplasmic translocation and PC interacts with MAVS and TRAF6.
(A) A549 cells were infected with SeV (MOI = 1) for 12 h or left uninfected. Mitochondrial extractions were performed and the fractions were analyzed by western blotting with the indicated antibodies. (B) 293T cells were infected with SeV (MOI = 1) for12 h or left uninfected. Western blotting was performed as in (A). (C) 293T cells were infected with SeV (MOI = 1) for12 h or left uninfected. Isolation of Mitochondria was then performed and the mitochondrial extracts were prepared by lysing mitochondrion in RIPA buffer. Co-immunoprecipitation and immunoblot analyses were performed with the MAVS, PC, Actin or COX4 antibodies. (D) A549 cells were transfected with PC or vector plasmids for 24 h. The cells were harvested after they were infected with SeV (MOI = 1) or left uninfected for 12 h. Co-immunoprecipitation and immunoblot analyses were performed with the MAVS, PC or GAPDH antibodies. (E) The experiment was performed as in (D) using the PC, TRAF6 or GAPDH antibodies. (F) Confocal immunofluorescence microscopy of PC and MAVS in A549 cells. MAVS/PC and nuclei were stained with FITC/Cy3-conjugated secondary antibodies and DAPI, respectively. (G) A549 cells were transfected with HA-TRAF6 for 48 h. HA/PC and nuclei were stained with Cy3/FITC-conjugated secondary antibodies and DAPI, respectively. Scale bars, 10 μm. All experiments were repeated at least three times with consistent results.
