Figure 3: FBXW7 positively regulates RIG-I signal pathway through protecting RIG-I from degradation.
From: E3 ligase FBXW7 is critical for RIG-I stabilization during antiviral responses
(a) Immunoblot analysis of phosphorylated or total proteins in lysates of FBXW7f/f and Lysm+FBXW7f/f peritoneal macrophages infected for indicated hours with VSV. (b) Immunoblot of RIG-I and TBK1 protein levels in lysates of FBXW7f/f and Lysm+FBXW7f/f macrophages infected with VSV. (c) Quantitative PCR (Q-PCR) analysis of RIG-I mRNA expression in FBXW7f/f and Lysm+FBXW7f/f macrophages infected for indicated hours with VSV. Data are mean±s.e.m. and are representative of three independent experiments. Student’s t-test was used for statistical calculation. NS, not significant. (d) Immunoblot analysis of RIG-I in lysates of FBXW7f/f and Lysm+FBXW7f/f peritoneal macrophages treated with CHX (40 μg ml−1) for indicated hours after infection with VSV for 1 h. (e) Quantification of relative RIG-I levels is shown in the right panel. (f) Confocal microscopy imaging of HEK293T cells that transfected with Flag-FBXW7, Myc-RIG-I for 24 h and then infected for indicated hours with VSV, and labelled with antibodies to the appropriate protein. Scale bar, 20 μm. (g) Coimmunoprecipitation and immunoblot of HEK293T cells transfected for 24 h with Flag-FBXW7 plasmid or Flag-FBXW7 together with Myc-RIG-I followed by VSV infection for indicated hours.
