Figure 2: USP13 restricts HSV-1-triggered signalling in THP-1 cells.
From: USP13 negatively regulates antiviral responses by deubiquitinating STING
(a) Immunoblot analysis (with anti-FLAG or anti-HA) of HEK293 cells transfected for 36 h with plasmids encoding FLAG-tagged USP13 and HA-β-Actin and either USP13-targeting shRNA (shUSP13#1 and shUSP13#2) or control shRNA (Con) to test efficiency of shRNA (upper panels). Immunoblot analysis (with anti-USP13 or anti-β-Actin) of THP-1 cells stably transfected with plasmids encoding control (Con) shUSP13 (#1 or #2) (lower panels). (b) qPCR analysis of IFNB, TNFA or IL6 mRNA in THP-1 cells stably transfected with shCon, shUSP13#1 or shUSP13#2 followed by infection with SeV or HSV-1 for 0–12 h. (c) Immunoblot analysis of phosphorylation of IRF3 and IκBα, ERK, JNK, or total IRF3 and IκBα, ERK, JNK, USP13 and β-Actin THP-1 cells stably transfected with shCon or shUSP13#1 followed by infection with HSV-1 for 0–12 h. **P<0.01; ***P<0.001 (analysis of two-way ANOVA followed by Bonferroni post-test). Data are representative of three independent experiments (mean±s.d. in b). See Supplementary Fig. 12 for uncropped immunoblots.
