Reciprocal negative regulation of miR-130a and HOTAIR. (A, B) Effect of miR-130a on HOTAIR expression. GBC-SD cells were transfected with vector, miR-130a mimic (A) or miR-130a inhibitor (B), and total RNA was isolated for qRT-PCR 24 h after transfection. (C) Schematic representation of the constructs generated for luciferase assays. (D) HOTAIR WT or its mutant devoid of specific miR-130a-binding sites in which seed matches were mutagenized from ‘TTGTAACGTGA’ to ‘GCGCCUCUUC’ was cloned downstream of Renilla luciferase gene (RLuc) in the vector pRL-TK and transfected into 293 T cells together with specific miRNAs mimics or the negative control mimic (NC). Luciferase assay was performed as described in Materials and Methods. Plotted are results from three independent experiments. Error bars represent S.E.M., n = 3. *p < 0.05; **p < 0.01, n.s., not significant.