Fig. 4: Oncogenic GAS5 sponges miR-423-3p for increased SMARCA4 ceRNA stability in various HCC cell lines.

a The SNU-182 or PLC/PRF/5 HCC cells were transfected with the psiCheck2-GAS5 vector and sequentially transfected with the indicated miRNAs. Then, the luciferase activity was measured with dual luciferase assays. b The relative expression of SMARCA4 was measured with qRT–PCR analysis (top) or western blotting analysis (bottom) in HCC cells, after they were transfected with si-Cont, si-GAS5, AS-miR-423-3p or AS-miR-452-5p. c A schematic presentation, the 5′-region of GAS5 and 3′UTR of SMARCA4 have miR-423-3p binding sites. The indicated sequences of either GAS5 (top) and SMARCA4 (bottom) each wild-type (WT) or mutant type (MUT) were inserted in the psiCHECK-2 plasmid. d psiCHECK-2 WT or MUT plasmids were transfected into SNU-182 or PLC/PRF/5 HCC cell lines, sequentially transfected with miR-423-3p and measured via dual luciferase activity. e The HCC cells were transfected with biotin-labeled microRNA control or biotin-labeled miR-423-3p mimics. The enrichment of GAS5 and SMARCA4 were measured by qRT–PCR, after pulldown with the biotin-labeled miR-423-3p. All the data are shown as the mean ± SEM, **P < 0.01, ***P < 0.001; unpaired t-test.