Fig. 6
LGP2 positively regulates fish IFN response dependently of MDA5
(A–C) LGP2-directed nuclear accumulation of IRF3 and IRF7 was significantly inhibited by functional blockade of MDA5 signaling. EPC cells seeded in 10-cm dishes were cotransfected with LGP2-HA at a low dose of 250 ng (left panels) or a high dose of 5 μg (right panels), together with the indicated dominant negative mutants (5 μg each). Forty-eight hours later, the transfected cells were harvested for fractionation of the nucleus and cytoplasm. IRF3, IRF7, and LGP2 proteins were detected by western blot analysis of whole-cell lysates as input (A), nuclear extracts (B), and cytoplasmic extracts (C), with antibodies specific to fish IRF3, IRF7, and LGP2, respectively. The red boxes highlighted the essential effects of MDA5-DN on LGP2-triggered signaling.