It was performed as described earlier [22 (link), 25 (link)]. Briefly, cells plated at 50–60% confluence in 12-well plates were co-transfected with 0.25 μg of tk-PPREx3-Luc, a PPRE-dependent luciferase reporter construct, and 12.5 ng of pRL-TK (a plasmid encoding Renilla luciferase, used as transfection efficiency control; Promega) using Lipofectamine Plus (Invitrogen Life Technologies). After 24 h of transfection, cells were stimulated with different doses of gemfibrozil for 2 h. Firefly and Renilla luciferase activities were analyzed in cell extracts using the Dual Luciferase kit (Promega) in a GloMax 96 microplate luminometer (Promega) and corresponding GloMax 96 microplate luminometer software (Promega) per the manufacturer’s specifications.
Protocol detail
Transcriptional Regulation of PPAR Targets
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Lipofectamine Plus
Dual Luciferase kit
GloMax 96 microplate luminometer
GloMax 96 microplate luminometer software
Cell extracts
Cells
Firefly
Gemfibrozil
Lipofectamine
Luciferase
Plasmid
Promega
Renilla luciferase
Transfection
Corresponding Organization :
Other organizations : Rush University Medical Center
Top 5 similar protocols
Protocol cited in 1 other protocol
Variable analysis
independent variables
- Different doses of gemfibrozil
- Firefly luciferase activity
- Renilla luciferase activity
- Cells plated at 50–60% confluence in 12-well plates
- Co-transfection with 0.25 μg of tk-PPREx3-Luc and 12.5 ng of pRL-TK using Lipofectamine Plus
- Transfection duration of 24 h
- Positive control: pRL-TK (a plasmid encoding Renilla luciferase, used as transfection efficiency control)
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.
Sign up for free or login to display all annotations
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!