Figure 5: FBXW7 interacts with SHP2.
From: E3 ligase FBXW7 is critical for RIG-I stabilization during antiviral responses
(a) Coimmunoprecipitation and immunoblot analysis of HEK293T cells transfected for 24 h with Flag-FBXW7 cells infected with VSV and treated with MG132. (b) Coimmunoprecipitation and immunoblot analysis of HEK293T cells cotransfected for 24 h with Flag-SHP2 plus HA-FBXW7 or Flag-FBXW7 and Myc-SHP2, and treated with MG132 followed by immunoprecipitation with anti-Flag-M2 beads. (c) Immunoblot analysis of RAW264.7 infected for indicate hours with VSV and treated with MG132, followed by immunoprecipitation with SHP2-conjugated agarose or immunoglobulin G (IgG)-conjugated agarose. (d) Confocal microscopy imaging of peritoneal macrophages infected for 3 h with VSV and labelled with antibodies to the appropriate protein. Scale bar, 20 μm. (e) Immunoblot analysis of HEK293T cells cotransfected for 24 h with Flag-FBXW7 or Flag-WD40, plus Myc-SHP2 vectors and treated with MG132, followed by immunoprecipitation with anti-Flag-M2 beads. (f) Immunoblot analysis of HEK293T cells cotransfected for 48 h with Flag-FBXW7, plus Myc-SHP2, Myc-SHP2 mutants vectors and treated with MG132, followed by immunoprecipitation with anti-Flag-M2 beads. *IgG heavy chain. (g) Sequence alignment of SHP2 with FBXW7 degron motif (〈RK〉S/TP〈RK〉XS/T/E/D), where X is any amino acid and 〈RK〉 is any amino acid, except arginine (R) or lysine (K). (h) Coimmunoprecipitation and immunoblot analysis of HEK293T cells transfected with Flag-FBXW7 along with vector for HA-SHP2 (WT) and HA-SHP2 (558A and 562A), and treated with MG132, followed by immunoprecipitation with anti-Flag-M2 beads.
