HepG2 cells were seeded in a 24-well culture plate. Plasmids of pGL3 basic or pGL3 basic-ARHGEF16 (0.75 µg) and pUB6/V5-HisB-Gli1 (0.25 µg), mixed with 0.025 µg pRL-TK-luc, were co-transfected into the cells using Lipofectamine™ 3000 transfection reagent (L3000015, Thermo Fisher Scientific, Inc.). The ARHGEF16 gene is a downstream target of Gli2 (8 (link)). This was conducted in triplicate, according to the manufacturer's instructions. After a 48-h transfection, the cells were treated with different concentrations of CH for 6 h. Cell lysates were extracted, and luciferase activity was measured using the Luciferase Report Assay System (E1910, Promega, Corp.).
Protocol detail
Luciferase Assay for ARHGEF16 Regulation
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.
Corresponding Organization :
Other organizations : Jinan University, Qinghai University Affiliated Hospital, Guangzhou Institutes of Biomedicine and Health
Top 5 similar protocols
Variable analysis
independent variables
- Concentration of CH
- Luciferase activity
- Cell line (HepG2)
- Plasmid constructs (pGL3 basic, pGL3 basic-ARHGEF16, pUB6/V5-HisB-Gli1, pRL-TK-luc)
- Transfection reagent (Lipofectamine™ 3000)
- Transfection duration (48 h)
- Treatment duration (6 h)
- Positive control: pGL3 basic-ARHGEF16 + pUB6/V5-HisB-Gli1
- Negative control: pGL3 basic + pUB6/V5-HisB-Gli1
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!