Fig. 9

Validation of miRNA binding to 3’ UTR of putative target genes. Luciferase reporter assay of HEK293 cells cotransfected with pmirGLO plasmid containing the WT or MT 3’UTR miRNA seed sequence from each gene, and either miR-29b, miR-223 or miR-NC (scrambled control). Samples were analysed 48 h after transfection and data normalised to the pmirGLO only transfection. Columns represent the luciferase activity of either WT or MT constructs with miR-29b (a) or miR-223 (b), relative to transfection with the same construct and miR-NC. Data represents the mean ± SEM, n = 3 independent experiments containing 4 replicates each. Student’s t test comparing WT or MT construct with miRNA to miR-NC indicated as *p < 0.05, **p < 0.01, ***p < 0.001. Student’s t test comparing WT with miRNA to corresponding MT construct indicated as #p < 0.05, ## p < 0.01, ###p < 0.001