Fig. 3. Experimental validation of miRNA-152-3p (miR-152) as a target gene of OIP5-AS1. (A) The putative binding site (in red above) of miR-152 in OIP5-AS1 was predicted by computational predictions on LncBase V.2 database. The potential target site in wild type of OIP5-AS1 (OIP5-AS1-WT) was mutated and shown in red below. (B) Luciferase reporter assay determined the relative luciferase activity of vectors containing OIP5-AS1-WT and its mutant (OIP5-AS1-MUT) when co-transfected with miR-152 mimic (miR-152) or miR-control mimic (miR-con) in hVSMCs. (C) RNA immunoprecipitation identified the enrichment of OIP5-AS1 and miR-152 in hVSMCs by Ago2 antibody and IgG antibody (negative control). (D and E) RT-qPCR detected effect of ox-LDL on miR-152 expression in hVSMCs when treated with gradient concentrations (0–200 μg mL⁻¹) of ox-LDL for 24 h, and 100 μg mL⁻¹ of ox-LDL for 0–48 h. (F) RT-qPCR analyzed miR-152 expression level in ox-LDL-treated hVSMCs transfected with pcDNA-OIP5-AS1 (OIP5-AS1), si-OIP5-AS1 and their negative controls. Data represent mean ± SEM. **P < 0.01 and ***P < 0.001 according to two-tailed Student's t-test.