Fig. 4

miR-34a directly interacts with MET mRNA and downregulates MET. a RNA was isolated from cells, biotin-based pull-down probe for miR-34a-5p or negative control probe at different concentrations to assess interaction of miR-34a-5p with MET in CAL27 cells was used. The enrichment of MET was quantified by qRT-PCR. b and c The miR-34a-5p overexpressing CAL27 cells were subjected to an anti-Ago2 CHIP assay. Levels of MET and miR-34a-5p were quantified by RIP-qPCR. d Dual-luciferase assay was performed in cells transfected with miR-34a-5p mimic or a control miR mimic and a luciferase reporter construct encoding the luciferase gene fused either to a mutated MET 3′ UTR (MUT) or a wild-type MET 3′ UTR (WT). e and f MET expression and miR-34a-5p expression in HNSCC cell lines were quantified by a TaqMan microRNA assay. g miR-34a-5p mimic was introduced to CAL27 cells by electroporation (25 nM). Levels of miR-34a-5p was quantified by a TaqMan microRNA assay after 12 h. h MET mRNA expression was quantified 24 h after over-expression of miR-34a-5p mimic. i Western blot measuring MET protein expression with and without expression of miR-34a-5p mimic. All experiments were performed in CAL27 cells. All experiments were performed in triplicate. Data represent results of at least three independent experiments. Data is presented as mean ± SD.* indicates p < 0.05