Fig. 6: SHP2 inactivation by LECT2 is essential for LECT2-mediated RIG-I dependent innate immune activation.

a HepG2 cells were treated with r-LECT2. Immunoblot analysis of p-SHP2, SHP2, p-GAB1, GAB1 and β-actin in r-LECT2-treated HepG2 cells. b Relative intensity. c Immunoblot analysis of K48-ubiquitin (Ub), RIG-I (FLAG), LECT2 (Myc) and β-actin in immunoprecipitated samples and input cell lysate. d qRT-PCR analysis of IFNB1 in HepG2-WT and HepG2-SHP2-KO cells at 12 h after Cont or Poly(I:C) transfection in the presence or absence of r-LECT2. Results were normalised to those of ACTB. Data shown are means of 3 technical replicates ± SEM. ***p < 0.001, not significant (ns) by two-way ANOVA with Tukey’s multiple comparison test. e Immunoblot analysis of SHP2, RIG-I, MDA5, p-IRF3, IRF3 and β-actin in HepG2-WT and HepG2-SHP2-KO cells at 12 h after Cont or Poly(I:C) transfection in the presence or absence of r-LECT2. f Immunoblot analysis of RIG-I, MDA5, p-IRF3, IRF3, p-ERK, ERK1/2 and β-actin in DMSO-treated HepG2 cells and SHP099-treated HepG2 cells at 12 h after HCV-RNA transfection in the presence or absence of r-LECT2. g qRT-PCR analysis of IFNB1, DDX58, IFIH1, OAS2 and MX1 in DMSO-treated HepG2 cells and SHP099-treated HepG2 cells at 12 h after HCV-RNA transfection in the presence or absence of r-LECT2. Data shown are means of 3 technical replicates ± SEM. ***p < 0.001, not significant (ns) by two-way ANOVA with Tukey’s multiple comparison test. h Immunoblot analysis of K48-Ub, SHP2, LECT2 (FLAG), RIG-I and β-actin in immunoprecipitated samples and input cell lysate. i HepG2-RIG-I-KO cells were transfected with RIG-I plasmid, SHP2 plasmid and FLAG-tagged LECT2 plasmid. At 48 h after transfection, the cells were transfected with HCV-RNA. qRT-PCR analysis of IFNB1 in these cells at 12 h after HCV-RNA transfection. Data shown are means of 3 technical replicates ± SEM. ***p < 0.001, **p = 0.001 by two-way ANOVA with Tukey’s multiple comparison test. Source data are provided as a Source data file.