Fig. 3

MET as a potential target of miR-199a-3p in WM-266-4 cells. a Relative expression of miR-199a-3p, DNMT1, EZH2, MET and COX2 in non-metastatic WM-115 and metastatic WM-266-4 GFP cell lines. Results are the mean of quadruplicates. MET mRNA and protein levels were measured upon miR-199a-3p ectopic expression (b) and 5azadC treatment (c). “Tox” indicates more than 50% of cytotoxicity. Histograms for mRNA expression are the mean of at least triplicates normalised to their respective control condition. For Western blots, one representative experiment out of three is shown. Densitometric measurements of MET expression were normalised to actin and relative to the control condition. d MET mRNA expression was monitored in WM-266-4 GFP cells, transfected with a siRNA targeting MET (siMET) at the indicated concentrations, by RT-qPCR (left panel) before testing cells for 3D invasion (right panel). The mean invasion index is calculated as previously and represented as histograms. SEM are shown. *P value < 0.05, **P value < 0.01, ***P value < 0.001. Ctl control and NT not treated