Fig. 3

MiR-218 directly targets IL-6R and JAK3 in lung cancer cells. a IL-6R and JAK3 gene was predicted as a direct target of miR-218 (TargetScan). b Relative luciferase activity was measured after luciferase reporter plasmids with IL-6R or JAK3 3′-UTR constructs (either wild-type or mutant) or control reporter plasmid were co-transfected into H1795 cells with miR-218 or miR-control, respectively. The data were the means ± SD of three individual experiments. c The mRNA expression of IL-6R and JAK3 was analyzed by quantitative real-time PCR 24 h after transfection of miR-218 or miR-control. d Protein levels of JAK3, IL-6R, pSTAT3 and STAT3 in A549 and H1795 cells were examined by western blotting 36 h after transfection of miR-218 or miR-control. e The phosphorylated STAT3 in H1975 and A549 cells was determined by immunofluorescence 36 h after transfection of miR-218 or miR-control. f Cell proliferation was determined by cell counting using H1975 and A549 cells 48 h after transfection of miR-218 or miR-control with or without IL-6 treatment