Fig. 1: The interaction of MEG3 with miR-21

a The prediction for MEG3 transcript binding sites on miR-21 by bioinformatics analysis. The nucleotides in bold are the complementary sequences to miR-21 seed sequences. b MEG3 is physically associated with miR-21. Curve lines (left) summarize the relative luciferase reporter activities with the increase of miR-21 concentration. Bar graph (right) shows relative luciferase activities in N2a cells cotransfected with miR-21 mimic (50 nM) + Vector, or miR-21 mimic (50 nM) + MEG3-WT, or miR-21 mimic (50 nM) + MEG3-MUT, or mimic control (50 nM) + Vector, or mimic control (50 nM) + MEG3-WT, or mimic control (50 nM) + MEG3-MUT. Data are means ± S.E.M. for 3 independent experiments. ^* P < 0.05 by Student’s t-test. Vector, luciferase reporters containing nothing; MEG3-WT, luciferase reporters containing the wild type MEG3; MEG3-MUT, luciferase reporters containing the mutant MEG3 with mutations of all three predicted miR-21-binding sites. c RNA pulldown assay based on MS2-MBP followed by miRNA qRT-PCR to detect miR-21 endogenously associated with MEG3. The schematic diagram on the left shows the strategies of the experiment. Bar graph (right) summarizes the relative expression of miR-21 in cells transfected with MEG3-WT-MS2 (MEG3-WT), MEG3-MUT-MS2 (MEG3-MUT) or the empty vector (MS2). Data are means ± S.E.M. for 3 independent experiments. ^** P < 0.01 by Student’s t-test