Figure 7: The role of FBXW7 stabilizing RIG-I and promoting the production of type I IFN is mediated by SHP2.
From: E3 ligase FBXW7 is critical for RIG-I stabilization during antiviral responses
(a) Quantitative PCR (Q-PCR) analysis of IFN-β and VSV-G mRNA expression in SHP2f/f and Lysm+SHP2f/f peritoneal macrophages infected with VSV. (b) Q-PCR analysis of IFN-β mRNA expression in SHP2f/f and Lysm+SHP2f/f peritoneal macrophages transfected for 24 h with Flag-FBXW7 and infected with VSV. (c) Q-PCR analysis of IFN-β mRNA in FBXW7f/f and Lysm+FBXW7f/f peritoneal macrophages transfected for 36 h with siSHP2 and infected with VSV. (d) Immunoprecipitation and immunoblot analysis of RAW264.7 infected for indicates hours with VSV followed by immunoprecipitation with FBXW7-conjugated magnetic bead or immunoglobulin G (IgG)-conjugated magnetic bead. (e) Immunoprecipitation and immunoblot analysis of FBXW7f/f and Lysm+FBXW7f/f peritoneal macrophages infected for indicate hours with VSV followed by immunoprecipitation with RIG-I-conjugated magnetic bead or immunoglobulin G (IgG)-conjugated magnetic bead. The equal amount of RIG-I for different groups was used for western blot analysis. (f) Immunoprecipitation and immunoblot analysis of SHP2f/f and Lysm+SHP2f/f peritoneal macrophages infected for indicated hours with VSV followed by immunoprecipitation with RIG-I-conjugated magnetic bead or immunoglobulin G (IgG)-conjugated magnetic bead. (g) Immunoblot analysis of RIG-I protein level in lysates of SHP2f/f and Lysm+SHP2f/f macrophages infected with VSV. (h) Immunoblot analysis of RIG-I protein in lysates of FBXW7f/f and Lysm+FBXW7f/f macrophages transfected for 36 h with SHP2-siRNA (siSHP2) and infected with VSV. Data are mean±s.e.m. and are representative of three independent experiments. Student’s t-test was used for statistical calculation. *P<0.05, **P<0.01 and ***P<0.001.
