Cloning EPAS1-derived Enhancer Assay

DNA fragments containing the EPAS1-derived EREs were generated by PCR using the primers listed in Supplementary Table 1B, and cloned into the pGL3-promoter luciferase reporter vector (Promega Corporation, Madison, WI, USA). A cDNA encoding human ERα cloned into the pCMV5 mammalian expression vector was kindly provided by A. Odermatt (Basel, Switzerland). Human GATA-2, GATA-3, and GATA-4 cloned into pcDNA3.1 were kindly provided by C. Dame (Berlin, Germany). Dual luciferase reporter gene assays were performed as described previously [64 (link)].

Free full text: Click here

Fuady J.H., Gutsche K., Santambrogio S., Varga Z., Hoogewijs D, & Wenger R.H. (2016). Estrogen-dependent downregulation of hypoxia-inducible factor (HIF)-2α in invasive breast cancer cells. Oncotarget, 7(21), 31153-31165.

Publication 2016

pGL3-promoter luciferase reporter vector

Assays Cdna Dame Epas1 Gata 3 Human Luciferase Mammalian Pgl3 Primers Promega Reporter gene Vector

Corresponding Organization : University of Zurich

Other organizations : University Hospital of Zurich, University of Duisburg-Essen

Top 5 similar protocols

Variable analysis

independent variables

DNA fragments containing the EPAS1-derived EREs

dependent variables

Luciferase reporter gene activity

control variables

PGL3-promoter luciferase reporter vector (Promega Corporation, Madison, WI, USA)
CDNA encoding human ERα cloned into the pCMV5 mammalian expression vector
Human GATA-2, GATA-3, and GATA-4 cloned into pcDNA3.1

controls

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

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!