USP49 interacts with MITA and inhibits MITA-mediated signaling.

(A) Immunoprecipitation (with anti-MITA) and immunoblot analysis (with anti-MITA or anti-USP49) of THP-1 or U937 cells infected with HSV-1, or transfected with ISD (10 μg) or cGAMP (4 μg) for the indicated time points. (B) Immunoprecipitation (with anti-Flag) and immunoblot analysis (with anti-FLAG or anti-HA) of HEK293 cells transfected with plasmids encoding HA-MITA and FLAG-tagged USP49 or USP49 truncates for 24 h or transfected with plasmids encoding FLAG-USP49 and HA-MITA or MITA truncates for 24 h. (C) qRT-PCR analysis of IFNB, IFNA4, CCL5 and IL6 in control and USP49 KO THP-1 cells infected with HSV-1 or SeV for 0–6 h. (D) Immunoblot analysis of phosphorylated and total IRF3, IκBα, TBK1 and Tubulin in control and USP49 KO THP-1 cells infected with HSV-1 or SeV for 0–6 h. *P P P <0.001 (analysis of two-way ANOVA followed by Bonferroni post-test). Data are representative of three independent experiments (mean ± S.D. in C).