Fig. 4

miR-16-5p was a target of AGAP2-AS1 in HCC. a Bioinformatics analysis showed that miR-16-5p could directly target 3′-UTR of AGAP2-AS1-wild type (WT). AGAP2-AS1-mutant (Mut) means mutation of binding sites in the 3′-UTR of AGAP2-AS1. b The expression of miR-16-5p in tumor tissues was significantly lower than that in adjacent nontumor tissues. c Pearson correlation analysis revealed that an obvious negative association between miR-16-5p and AGAP2-AS1 expression in HCC tissues. d Real-time PCR showed that AGAP2-AS1 could negatively regulate miR-16-5p expression in HCC cells. e Dual luciferase reporter assays showed that miR-16-5p could negatively regulate the luciferase activity of AGAP2-AS1-WT, rather than AGAP2-AS1-Mut. f The association between AGAP2-AS1, miR-16-5p and Ago2 was ascertained by analyzing Hep-3B cell lysates using RNA immunoprecipitation with an Ago2 antibody. Real-time PCR was used to detect the AGAP2-AS1 level change in the substrate of RIP assay in miR-16-5p-overexpressing HCC cells. g Detection of AGAP2-AS1 using real-time PCR in the sample pulled down by biotinylated AGAP2-AS1 and negative control (NC) probe. Detection of miR-16-5p using real-time PCR in the same sample pulled down by biotinylated AGAP2-AS1 and NC probe. h miR-16-5p inversely regulate AGAP2-AS1 expression in HCC cells. *P < 0.05, ***P < 0.001