Figure 1.

LncRNA CR749391 directly interacts with miR-181a. (A) CR749391 was identified as a novel lncRNA located upstream of KLF6. (B) The 4 putative binding sites of miR-181a in the CR749391 transcript sequence. (C) Luciferase reporter plasmid containing the sequence from the 3′-untranslated region of KLF6 was co-transfected with miR-181a and CR749391-coding plasmids into 293T cells. Luciferase activity was determined at 48 hours after transfection using the dual luciferase assay and the results are presented in the histogram as the relative luciferase activity normalized to Renilla activity. *P<0.05, **P<0.01 vs. the control group. (D) RNA immunoprecipitation with mouse monoclonal anti-Ago2, pre-immune IgG or 10% input from BGC 823-cell extracts. RNA levels of miR-181a and CR749391 in immunoprecipitates were determined by reverse transcription-quantitative polymerase chain reaction. ^#P<0.05, ^##P<0.01 vs. Anti-IgG. (E) Pull-down analysis of CR749391 levels. ^&P<0.05 vs. Control. Values are expressed as the mean ± standard deviation (n=3). lncRNA, long non-coding RNA; miR, microRNA; chr, chromosome; mut, mutated; UTR, untranslated region; KLF, Kruppel-like factor; NC, negative control; IgG, immunoglobulin G; Ago2, argonaute 2, RISC catalytic component; LOC, locus.