Hypoxia-Responsive Luciferase Assay

DLK-S expression plasmid was kindly provided by prof. Anne Ferguson-Smith [11] (link), cloned into RCAS vector by classic restriction enzyme technique and transfected into DF-1 cells. For luciferase reporter assay, cells were co-transfected with hypoxia-responsive element (HRE)-luc (Addgene) [26] (link) or 8xCSL-luc (gift from Håkan Axelson) and pCMV-renilla (Promega) and analyzed using the Dual-Luciferase Reporter Assay System (Promega) on a Synergy 2 platereader (BioTek). Xtreme gene 9 (Roche) reagent was used according to manufacturer's recommendations for transient transfections.

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Grassi E.S., Jeannot P., Pantazopoulou V., Berg T.J, & Pietras A. (2020). Niche-derived soluble DLK1 promotes glioma growth. Neoplasia (New York, N.Y.), 22(12), 689-701.

Publication 2020

HRE)-luc pCMV-renilla Dual-Luciferase Reporter Assay System Synergy 2 platereader Xtreme gene 9

Assay Cells Co element Gene Hypoxia Luciferase Plasmid Promega Renilla Restriction enzyme Transfections Transient Vector

Corresponding Organization : Lund University

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Variable analysis

independent variables

Transfection of DLK-S expression plasmid into DF-1 cells
Co-transfection of cells with hypoxia-responsive element (HRE)-luc or 8xCSL-luc and pCMV-renilla

dependent variables

Luciferase reporter activity measured using the Dual-Luciferase Reporter Assay System

control variables

Use of Xtreme gene 9 (Roche) reagent for transient transfections according to manufacturer's recommendations

positive controls

Hypoxia-responsive element (HRE)-luc (Addgene)
8xCSL-luc (gift from Håkan Axelson)

negative controls

Not explicitly mentioned

Annotations

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