Fig. 3

Flavivirus TBEV prM interacts with MDA5 and MAVS. A HEK293T cells transfected with TBEV prM plasmid were fixed and stained by Calnexin (endoplasmic reticulum), GM130 (Golgi), COIXV (mitochondria), and Flag antibodies to analyze the location of prM. Green, the corresponding organelles signal; Red, TBEV prM signal; Blue, DAPI (the nuclear signal). Intensity profiles of the indicated proteins were analyzed by Image J line scan analysis. Bar, 10 μm. B–F EV or TBEV prM alone (F) or together with HA-MDA5 (B, D), Myc-MAVS (C, E) were transfected into HEK293T cells, cells were harvested at 30 hpt and the cell lysates were co-immunoprecipitated and analyzed by immunoblot using the indicated antibodies. G The diagram of TBEV prM truncations. TM, trans-membrane domain. Numbers above the domain names indicate amino acid positions of prM. H and I Co-immunoprecipitation and immunoblot analyses of the indicated proteins in HEK293T cells transfected with the full length (FL) and truncated fragments of TBEV prM along with MDA5 (H) or MAVS (I). J The expression plasmids of TBEV-prM and its truncations were co-transfected with an IFNβ-Luc and poly (I:C), cells were harvested for luciferase reporter assay at 20 hpt. Bars represent the mean of three biological replicates and all data are expressed as mean ± SE