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. 2013 Apr 3;41(9):4976–4987. doi: 10.1093/nar/gkt182

Figure 5.

Figure 5.

Reciprocal negative regulation of miR-211 and loc285194. (A) Loc285194 carries two miR-211 binding sites at exon 4. Alignment between them is shown underneath with miR-211 seed sequences underlined. (B) Suppression of miR-211 expression by loc285194. HCT-116 cells were first transfected with vector, wild-type loc285194 (Loc-WT) or mutant loc285194 (Loc-d) with a deletion of two miR-211 binding sites, and 24 h after transfection, total RNA was isolated for qRT-PCR. Error bars represent SEM, n = 3. **P < 0.01. (C) Effect of miR-211 on loc285194 expression. HCT-116 WT cells were transfected with vector or miR-211, and total RNA was isolated for qRT-PCR 24 h after transfection. Error bars represent SEM, n = 3. *P < 0.05. (D) Effect of loc285194 on mature miR-211, pri-miR-211 and pre-miR-211. Error bars represent SEM, n = 3. **P < 0.01; n.s., not significant. (E) Pulldown of Ago2 by biotin-labeled loc285194 or GAS5 RNA probe, as detected by western blot. The GAS5 lane was composed from the same gel with the same contrast. (F) Detection of miR-211 in the pellet precipitated by the loc285194 probe, but not in the pellet precipitated by the GAS5 probe. Error bars represent SEM, n = 3. **P < 0.01.