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. 2016 Jun 9;7(6):e2256. doi: 10.1038/cddis.2016.159

Figure 3.

Figure 3

Validation of miR-206 targets. (a) qRT-PCR and (b) immunoblot analysis of putative miR-206 targets 48 h after NC or miR-206 mimic transfection in RD cells using 5 μg (upper panels) or 25 μg (lower panels) of whole-cell lysate. All differences by qRT-PCR with exception of EZH1 Student's unpaired t-test P<0.05. (c) Luciferase activity in 293T cells co-transfected with miR-206 or control vector, and empty psiCHECK2 reporter, miR-206 sensor positive control and wild-type 3′ UTRs of indicated genes, including a non-targeting control (EZH1), P<0.0001 for all excluding EZH1. (d) Luciferase activity in 293T cells co-transfected with miR-206 or control vector and wild type or mutant miR-206 site responsive 3′ UTRs, P<0.0005 for all comparing WT and mutant reports with miR-206. Luciferase activity represented as mean±S.E.M. (n=4), the Renilla/Firefly luciferase ratio normalized to empty reporter (no miR-206)