Improve Research Reproducibility A Bio-protocol resource
Concise Method
tooltip

2.12. Dual-Luciferase Reporter Assay and Transfections

ZP Zhengjun Peng
LL Lu Liu
WZ Wenyu Zhang
XW Xi Wei
request Request a Protocol
ask Ask a question
Favorite

The target genes for transcription factor STAT3 were identified by TargetScan. pGL3 plasmids (Promega, Madison, WI, USA) containing a STAT3 binding site for wild-type (WT) or MUT (mutated STAT3 binding site) Oct-4/Sox2 were amplified and then transfected along with STAT3 mimics into HEK293T cells. A control vector (pTK-RL) containing the Renilla luciferase gene was added to the transfection mix. Transfections were performed by using Lipofectamine 2000 (Invitrogen) according to the manufacturer's protocol. After 48 h of transfection, luciferase (Firefly/Renilla) activity was measured using the Dual-Glo™ Luciferase Assay System (Promega) as described in the manufacturer's protocol.

Copyright and License information:  ©2021 Zhengjun Peng et al.
This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A