miR-941 targets KDM6B and negatively regulates KDM6B at the post-transcriptional level.
A, the predicted binding sites for miR-941 on the 3′ UTR of KDM6B mRNA are shown. The mutant 3′ UTR contains 5 mutated nucleotides in the complementary seed sequences. B and C, relative EGFP activity was analyzed after the wild-type or mutant 3′ UTR reporter plasmids were cotransfected with miR-941 or ASO-miR-941 in QGY-7703 cells. The histogram shows the mean ± S.D. of the normalized EGFP intensity from three independent experiments. D, qRT-PCR shows the mRNA level of KDM6B after the cells were transfected with pri-miR-941 and ASO-miR-941 compared with the control group. E, KDM6B immunostaining (green) and DAPI staining (blue) in QGY-7703 cells after transfection. F, Western blot shows the protein level of KDM6B after the cells were transfected with pri-miR-941 and ASO-miR-941. G, different expression of KDM6B in 15 pairs of HCC tissues compared with adjacent normal tissues. H, different expression of KDM6B in six different HCC cell lines compared with the non-cancerous cell (LO2) (*, p < 0.05; **, p < 0.01).