Fig. 6

MiR-486-5p suppresses EVI5 expression and reduces the proliferation, migration and invasion capability of NSCLC cells. a Schematic diagram showing the subcloning of a fragment containing the predicted miR-486-5p-binding site at positions 765–770 in the EVI5 3′-UTR into the psiCHECK-2 luciferase vector. Predicted duplex formation between miR-486-5p and the wild-type or mutant miR-486-5p-binding site is indicated. b-c Luciferase activity of the constructs containing the wild-type or the mutant EVI5 reporter gene in A549 and H226 cells co-transfected with miR-NC or miR-486-5p. A scrambled sequence was used as the NC. The relative Renilla luciferase activity was measured and normalized to firefly luciferase activity. d-e The expression of miR-486-5p and EVI5 in NSCLC cells transfected with miR-486-5p mimics was measured by qRT-PCR. f CCK-8 assay of cell viability in EVI5-overexpressing A549 cells transfected with miR-486-5p mimics or miR-NC. g Representative images of the clonogenic assay results for cell proliferation in EVI5-overexpressing A549 cells transfected with miR-486-5p mimics or miR-NC. h Representative images of the transwell assay results for cell migration and invasion in EVI5-overexpressing A549 cells transfected with miR-486-5p mimics or miR-NC. i EVI5-overexpressing A549 cells were transfected with miR-486-5p or miR-NC, and the expression levels of various proteins were then measured by western blot analysis. j The mRNA expression levels of miR-486-5p were measured by qRT-PCR and compared between 26 NSCLC and paired adjacent noncancerous lung tissues. k qRT-PCR analysis of relative miR-486-5p expression levels in human NSCLC cell lines. β-actin was used as the internal control. Bars represent mean ± SD from three independent experiments. Significant differences compared with the control: * P < 0.05; ** P < 0.01; ***P < 0.001