Research Paper Volume 15, Issue 19 pp 10105—10116

Figure 5. SNHG25 acts as a sponge for miR-296-3p in CRC. (A) Relative expression of microRNAs which potentially bind to SNHG25 were detected by qRT-PCR after RNA pull-down assays in HCT-116 cells with biotin labeled SNHG25. (B) MiR-296-3p expression was detected by qRT-PCR in HCT-116 cells transfected with SNHG25 siRNAs. (C) MiR-296-3p expression was detected by qRT-PCR in HCT-116 cells transfected with pcDNA3.1-SNHG25. (D) RIP assays were performed with antibodies against AGO2 or control IgG in HCT-116 cells. Immunoprecipitated RNA was analyzed by qRT-PCR. Total RNA input was 1%. (E) Dual luciferase reporter assays with wild type and mutant (binding sites for miR-296-3p were mutated) SNHG25 luciferase reporters. Up panel, sequence alignment of miR-296-3p and its potential binding sites in SNHG25. Predicted miR-296-3p target sequence (blue) in Luc-SNHG25-Wt and mutated nucleotides (red) in Luc-SNHG25-Mut. (F) MiR-296-3p expression was detected by qRT-PCR in 90 pairs of CRC tissues and adjacent normal tissues. *P < 0.05, **P < 0.01 and ***P < 0.001.