Fig. 4: MEG3 competes with PDCD4 mRNA for binding to miR-21 | Cell Death & Disease

Fig. 4: MEG3 competes with PDCD4 mRNA for binding to miR-21

From: Long non-coding RNA MEG3 functions as a competing endogenous RNA to regulate ischemic neuronal death by targeting miR-21/PDCD4 signaling pathway

Fig. 4

a miR-21 bounds to 3’-UTR of PDCD4 mRNA by bioinformatics analysis. Overlapping sequences of the binding sites of MEG3–miR-21 and PDCD4–miR-21 are marked with a rectangular box. b The modulation of MEG3 levels by MEG3-expressing plasmids or si-MEG3 in N2a cells were confirmed by qRT-PCR. Data are means ± S.E.M. for 3 independent experiments. * P < 0.05, **P < 0.01, by Student’s t-test. c N2a cells were cotransfected with mimic control + Vector, or miR-21 + Vector, or mimic control + PDCD4-WT, or miR-21 + PDCD4-WT, or mimic control + PDCD4-MUT, or miR-21 + PDCD4-MUT. Relative luciferase activity was measured by luciferase reporter assays. Data are means ± S.E.M. for 3 independent experiments. * P < 0.05 by Student’s t-test. miR-21, miR-21 mimic; Vector, luciferase reporters containing nothing; PDCD4-WT, luciferase reporters containing wild type PDCD4; PDCD4-MUT, luciferase reporters containing mutant PDCD4. d N2a cells were cotransfected with si-s-MEG3 + Vector, or si-MEG3 + Vector, or si-s-MEG3 + PDCD4-WT, or si-MEG3 + PDCD4-WT, or si-s-MEG3 + PDCD4-MUT, or si-MEG3 + PDCD4-MUT. Relative luciferase activity was measured by luciferase reporter assays. Data are means ± S.E.M. for 3 independent experiments. * P < 0.05 by Student’s t-test. si-s-MEG3, negative control of scramble RNA for si-MEG3; si-MEG3, MEG3-targeting small interference RNAs. e N2a cells were cotransfected with pcDNA-vector + Vector, or MEG3 + Vector, or pcDNA-vector + PDCD4-WT, or MEG3 + PDCD4-WT, or pcDNA-vector + PDCD4-MUT, or MEG3 + PDCD4-MUT. Relative luciferase activity was measured by luciferase reporter assays. Data are means ± S.E.M. for 3 independent experiments. * P < 0.05 by Student’s t-test. pcDNA-vector, pcDNA3.0 empty vector which was used to MEG3-expressing plasmids construction; MEG3, MEG3-expressing plasmids; f N2a cells were cotransfected with PDCD4-WT, or miR-21 + PDCD4-MUT, or miR-21 + PDCD4-WT, or miR-21 + MEG3 + PDCD4-WT, or miR-21 + si-MEG3 + PDCD4-WT. Relative luciferase activity was measured by luciferase reporter assays. Data are means ± S.E.M. for 3 independent experiments. * P < 0.05 by Student’s t-test. NS, not significant

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