Fig. 8: PTP4A1 recruitment to MET is essential for LECT2-mediated RIG-I dependent innate immune responses.

a HepG2 cells were treated with r-LECT2 (0, 5, 10, 50 ng/mL). At 6 h after treatment, lysates were immunoprecipitated with an anti-MET antibody. Immunoblot analysis of MET, p-MET (Tyr1349), p-SHP2, SHP2, PTP4A1, c-Cbl and β-actin in immunoprecipitated samples and input cell lysate. b HepG2-MET-KO cells were transfected with Control plasmid (Cont), MET WT plasmid, MET Y1349F plasmid, or PTP4A1 plasmid. At 48 h after transfection, lysates were immunoprecipitated with an anti-MET antibody. Immunoblot analysis of MET, p-MET (Tyr1349), p-SHP2, SHP2, PTP4A1, c-Cbl and β-actin in immunoprecipitated samples and input cell lysate. c HepG2 cells were transfected with 50 nM c-Cbl siRNA. The concentration of siRNA was adjusted to 50 nM with control siRNA. At 24 h after siRNA transfection, siRNA-transfected HepG2 cells were treated with 50 ng/mL r-LECT2 for 1 h and then transfected with HCV-RNA. c (upper) Immunoblot analysis of c-Cbl, RIG-I and β-actin at 12 h after HCV-RNA transfection. c (lower) qRT-PCR analysis of IFNB1 at 12 h after HCV-RNA transfection. 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. d HepG2-MET-KO cells were transfected with Control plasmid or MET plasmid; at 48 h after transfection, the cells were treated with 50 ng/mL r-LECT2 and/or 10 ng/mL HGF. At 1 h after r-LECT2 and/or HGF treatment, lysates were immunoprecipitated with an anti-MET antibody. Immunoblot analysis of MET, p-MET (Tyr1234/1235), p-MET (Tyr1349), p-MET (Tyr1356), PTP4A1, p-SHP2, SHP2 and β-actin in immunoprecipitated samples and input cell lysate. Source data are provided as a Source data file.