Fig. 5.

Lnc-TALC competitively binds miR-20b-3p targeting the MET 3′UTR region. a Schematic process of the RIP-PCR assay. b RIP-PCR assay of the enrichment of Ago2 on lnc-TALC and c-MET transcript normalized to IgG in LN229 or 229R cells transfected with LV-scramble, or LV-lnc-TALC or LV-CRISPR-lnc-TALC (n = 4). c Schematic outline of predicted and mutant binding sites for miR-20b-3p on lnc-TALC and MET. d Luciferase activity of GV-272-lnc-TALC or GV-272-MET upon transfection of control mimics and miR-20b-3p in 229R (n = 3). e Upper: Schematic process of the MS2-RIP assay. Lower: MS2-based RIP assay with anti-GFP antibody in LN229 cells 72 h after transfection with MS2bs-Rluc, MS2bs-lnc-TALC or MS2bs-lnc-TALC-Mut (n = 4). f Upper: Luciferase activity of GV-272-MET in 229R lnc-TALC knockdown upon transfection of a miR-20b-3p sponge (n = 3). Lower: Luciferase activity of GV-272-MET in LN229 overexpressing lnc-TALC upon transfection of miR-20b-3p mimics (n = 3). g Western blot analysis of c-Met in 229R and HG7R cells transfected with LV-CRISPR-lnc-TALC or a miR-20b-3p sponge. h Western blot analysis of c-Met in LN229 and HG7 cells transfected with LV-lnc-TALC or miR-20b-3p mimics. Data are presented as the mean ± S.D. P value was determined by Student’s t-test or one-way ANOVA. Significant results were presented as NS non-significant, *P < 0.05, **P < 0.01, or ***P < 0.001