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. 2018 May 2;4(5):eaar2824. doi: 10.1126/sciadv.aar2824

Fig. 4. DUBs regulate type I IFN signaling by altering ubiquitination in both catalytic activity–dependent and –independent manners.

Fig. 4

(A) Heat map summary of regulatory roles of DUBs on total ubiquitination of key molecules in the type I IFN signaling pathway. (B) Heat map summary of regulatory roles of DUBs on ubiquitination of key molecules with different linkages in the type I IFN signaling pathway. Data are obtained by band intensity analysis (IP-HA/IP-Flag) and are normalized to the control sample. (C) Luciferase activity in 293T cells transfected with an IFN-β luciferase reporter, together with vectors for RIG-I (2CARD), along with an empty vector or with expression vectors for the indicated WT DUB or catalytically inactive mutants. (D) Coimmunoprecipitation and immunoassay of extracts of 293T cells transfected with Flag-TRAF3, Myc-BRCC3, and Myc-BRCC3-H122Q with the indicated antibodies. (E) Coimmunoprecipitation and immunoassay of extracts of 293T cells transfected with Flag-RIG-I and HA-K11-Ub together with Myc-USP5 or Myc-USP5-C335A followed by SeV infection with the indicated antibodies. (F) Coimmunoprecipitation and immunoassay of extracts of 293T cells transfected with Flag-STING and HA-K27-Ub together with Myc-USP22 or Myc-USP22-C185A followed by HSV-1 infection with the indicated antibodies. (G) Coimmunoprecipitation and immunoassay of extracts of 293T cells transfected with Flag-RIG-I and HA-K48-Ub together with Myc-USP5 or Myc-USP5-C335A with the indicated antibodies. Data are representative of three independent experiments [mean and SEM in (C)]. NS, nonsignificant (P > 0.05); ***P < 0.001.