Journal: Molecular Therapy. Nucleic Acids

Article Title: MEG3, as a Competing Endogenous RNA, Binds with miR-27a to Promote PHLPP2 Protein Translation and Impairs Bladder Cancer Invasion

doi: 10.1016/j.omtn.2019.01.014

Figure Lengend Snippet: c-Jun Was Essential for MEG3 Inhibition of c-Myc Transcription in Human Bladder Cancer Cells (A) Diagram of the potential transcription factor binding sites in human c-Myc promoter-driven luciferase reporter. (B) The effect of MEG3 overexpression on the expression of potential transcription factors listed in (A), with GAPDH used as the internal loading control. (C) The effect of MEG3 on AP-1 activity in UMUC3 cells. The AP-1 luciferase reporter was co-transfected into UMUC3(Vector) and UMUC3(MEG3) cells with TK at a ratio of 10:1. After 24 h, the luciferase activity was determined and presented as luciferase activity relative to vector control. (D and E) Ectopic expression of TAM67 on c-Myc expression at the protein (D) and the mRNA (E) levels in UMUC3 cells. GAPDH was used as the internal loading control. (F) Diagram indicating the c-Myc promoter-driven luciferase reporter Del2 and its c-Jun binding site point mutation. (G) WT and mutant (Mut) c-Myc promoter-driven luciferase reporters were stably co-transfected with MEG3 or its scrambled vector into T24T cells, and the stable transfectants were used to evaluate the essential role of the c-Myc binding site in MEG3’s inhibition of c-Myc promoter transcription. Results are presented as relative c-Myc promoter activity. The asterisk indicates a significant inhibition compared with the vector transfectant (p < 0.05). The bars represent the mean ± SD of triplicates.

Article Snippet: The c-Myc promoter luciferase reporter, HA-MS2-GFP overexpression vector and pSL-MS2-12X plasmids were obtained from Addgene (Cambridge, MA, USA).

Techniques: Inhibition, Binding Assay, Luciferase, Over Expression, Expressing, Activity Assay, Transfection, Plasmid Preparation, Mutagenesis, Stable Transfection