Fig. 4.

tMDA5 interacts with tMITA in TSPRCs. Overexpression tMDA5 (A) and tMITA (B) activate the IFN-β-Luc, NF-κB-Luc, and ISRE-Luc reporters in a dose-dependent manner. Cells (1 × 10⁴) were transfected with the respective reporter vector (100 ng), TK (10 ng), and increased amount (0, 3.2, 16, 80, and 400 ng) of tMDA5 or tMITA expression vector (with empty vector to reach a total amount of 400 ng) for 36 h and then were transfected with poly I:C (1 μg/mL) for 12 h. (C) Knockdown of tMITA causes an inhibition of poly I:C-induced IFN-β-Luc, NF-κB-Luc, and ISRE-Luc activities. (D) tMDA5 immunoprecipitates with tMITA. Cells (1 × 10⁷) were cotransfected with MYC-tMITA (10 μg) and Flag-tMDA5 (10 μg) expression vectors for 36 h and then were infected with SeV for 16 h, followed by immunoprecipitation (IP) with anti-Flag, anti-MYC, or mouse IgG (control). The EGFP-tagged tMAVS was used as a positive control. Data are representative of three independent experiments.*P < 0.05, **P < 0.01, ***P < 0.001, Student t test. Bars represent mean ± SEM.