Figure 3: USP13 deficiency potentiates DNA virus-triggered signalling.
From: USP13 negatively regulates antiviral responses by deubiquitinating STING
(a) qRT-PCR analysis of Ifnb, Ifna4, and Tnf mRNA in Usp13+/+ and Usp13m/m BMDCs left uninfected (Mock) or infected with HSV-1, SeV or EMCV for 3–6 h or mock transfected (Lipo) or transfected with ISD45, DNA90, HSV120 or poly(I:C) for 6 h. (b) ELISA analysis of IFN-α, TNFα and IL-6 in the supernatants of Usp13+/+ and Usp13m/m BMDC infected for 12–24 h, or mock transfected (Lipo) or transfected with ISD45, DNA90 or poly(I:C) for 12 h. (c) Immunoblot analysis of phosphorylation of IRF3 and IκBα, total IRF3 and IκBα, USP13 and β-Actin in Usp13+/+ and Usp13m/m BMDCs infected with SeV or HSV-1 for 0–8 h. (d) qPCR analysis of HSV-1 UL30 mRNA in Usp13+/+ and Usp13m/m BMDCs (5 × 105) left uninfected or infected with HSV-1 (MOI=0.1) for 1 h followed by twice PBS wash and cultured in full medium for 24 h (left graph). Plaque assay of HSV-1replication in the supernatants of Usp13+/+ and Usp13m/m BMDCs infected with HSV-1 (MOI=0.1) for 1 h followed by twice PBS wash and cultured in full medium for 36 h (right graph). *P<0.05; **P<0.01; ***P <0.001 (analysis of two-way ANOVA followed by Bonferroni post-test). PFU, plaque-forming unit. Data are representative of three independent experiments (mean±s.d. in a,b,d). See Supplementary Fig. 12 for uncropped immunoblots.
