Fig. 4

The lncRNA NEAT1 sponges miR-483 and suppresses its expression. a The putative binding site for miR-483 in the NEAT1 according to the bioinformatics prediction from the Starbase database. b U2OS and MG-63 cells were transfected with a luciferase plasmid containing the wild type or mutant 3′UTR of NEAT1 as along with miR-483 mimics or mimic NC. The interaction between miR-483 and the 3′UTR of NEAT1 was assessed using the dual luciferase assay. c U2OS and MG-63 cells were transfected with miR-483 mimics or mimic NC, and the interaction between NEAT1 and miR-483 was confirmed using an RIP assay. d U2OS and MG-63 cells were untransfected or transfected with a negative control vector (OE-NC) or NEAT1 overexpression plasmid (OE-NEAT1), and the relative expression of miR-483 was determined using qRT-PCR. The expression of miR-483 was normalized to U6. All experiments described in this study were performed at least three times, which referred to biological replicates. Each biological replicate consisted of 3 wells with cells of the same passage, which referred to technical replicates. Error bars denoted mean ± SD. p values were calculated using unpaired two-tailed Student’s t-test (b, c) or one-way analysis of variance (ANOVA) followed by Tukey’s post hoc test (d). **p < 0.01 and *p < 0.05