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. 2017 Jun 21;14(2):2325–2333. doi: 10.3892/ol.2017.6444

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Copyright: © Zhang et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Figure 2. — Notch1 and Jagged1 are direct target genes of human miR-34a. (A) Schematic representation of the predicted binding sites of miR-34a in the Notch1 and Jagged1 3′ untranslated regions, and the sequence of the intact (wild-type) miR-34a and their mutant binding sites within the luciferase reporter vector. (B) miR-34a suppresses the expression of Notch1 and Jagged. Reverse transcription-quantitative polymerase chain reaction (middle panel) and Western blot analyses (right panel) of Notch1 and Jagged1 were performed following transfection of cells with an miR-34a mimic or mimic negative control (NC) for 48 h (left panel). The data are expressed as the mean ± SD (n=3). *P<0.05 and **P<0.01 vs. mimic NC. (C) A total of 9 luciferase reporter constructs were created for the luciferase activity assay, namely LR-blank (no insertion), LR-34a/Noth1w1, LR-34a/Noth1w2, LR-34a/Jagged1w1, LR-34a/Jagged1w2, LR-34a/Noth1m1, LR-34a/Noth1m2, LR-34a/Jagged1m2 and LR-34a/Jagged1m2. Each of the constructs was co-transfected with an miR-34a mimic (or mimic NC) into HEK293 cells. Luciferase activities were measured 36 h post-transfection. The data are expressed as the mean ± SD (n=3). *P<0.05 and **P<0.01 vs. mimic NC. miR, microRNA; CDS, coding sequence; SD, standard deviation; NC, negative control; w1/2, wild-type 1/2; m1/m2, mutant 1/2.