Figure 5.

H-Ras and H-RasN17 enhances the formation of the MAVS–TNF receptor-associated factor 3 (TRAF3) signaling complex, whereas H-RasV12 reduces it. (A) HEK293T cells were depleted of H-Ras by shHRAS#64 or #66, or (B) cells were transfected with vector, the HRASV12 DNA (15 ng/ml), or the HRASN17 DNA (60 ng/ml) at the indicated doses shown in the parenthesis. In all groups, the total amounts of transfected DNA were adjusted to the same using the control vector DNA. After knockdown or DNA transfection, the cells were then mock infected or infected with Sendai virus (SeV) (20 HAU/ml) for 24 h. Cell lysates were immunoprecipitated with an anti-TRAF3 antibody or control IgG antibody, followed by Western blot analysis using anti-MAVS and anti-TRAF3 antibodies. WCL, whole cell lysate. The ratios of immunoprecipitated MAVS relative to total MAVS are shown at the bottom.