Validating miR-1956 Target Genes via Luciferase Assay

To validate miR-1956 target genes, we performed luciferase reporter assays on HEK293 cells as previously described 21 (link). Briefly, cells were transiently transfected using TransIT-LT1 (Mirus) according to the manufacturer's protocol. A dual-luciferase reporter assay system (Promega) and a Sirius single tube luminometer (Berthold) were used to determine luciferase activity. The Renilla luciferase coding vector (Promega) was co-transfected in all cases to evaluate transfection efficiency. Luciferase experiments were performed in quadruplicate and repeated three times independently. Luciferase activity results are presented as relative light units (RLU): the average of Photinus pyralis luciferase activity was divided by the average of Renilla luciferase activity.

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Gao L., Mei S., Zhang S., Qin Q., Li H., Liao Y., Fan H., Liu Z, & Zhu H. (2020). Cardio-renal Exosomes in Myocardial Infarction Serum Regulate Proangiogenic Paracrine Signaling in Adipose Mesenchymal Stem Cells. Theranostics, 10(3), 1060-1073.

Publication 2020

TransIT-LT1 dual-luciferase reporter assay system Sirius single tube luminometer

Assay Cells Genes Hek293 cells Light Luciferase Photinus luciferase Promega Renilla luciferase Transfection Vector

Corresponding Organization : Shanghai East Hospital

Other organizations : Renji Hospital, Shanghai Jiao Tong University, Fudan University, Zhongshan Hospital, Shanghai Tenth People's Hospital, Tongji University

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Variable analysis

independent variables

Transfection with miR-1956 target gene constructs

dependent variables

Luciferase activity

control variables

Transfection efficiency (measured by co-transfection of Renilla luciferase coding vector)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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