RORβ-overexpression GC cells were seeded into 6-well plates at a density of 1×106 at 37°C with 5% CO2 for 24 h. A total of 8 µl Fugene 6 (Roche Diagnostics) was incubated with 100 µl OPTI-MEM (Invitrogen; Thermo Fisher Scientific, Inc.) at room temperature for 5 min. Then, a total of 2 µg plasmid mixture including TOP (MilliporeSigma), FOP (MilliporeSigma) and PRL (Promega Corporation) (4:4:1) were added to the aforementioned mixture and incubated at room temperature for 15 min. Then the mixture of Fugene 6 and plasmids were added to the 6-well plate and mixed, and incubated at 37°C in a 5% CO2 incubator for 24 h. The cell lysates were transferred to the centrifuge tube after cell lysis. GloMax 20/20 Luminometer was used to detect the fluorescence activity. A total of 20 µl of the aforementioned cell lysates were added with 100 µl of LAR II. After mixing, firefly luciferase activity was detected. Then 100 µl Stop&Glo Reagent was added to detect Renilla luciferase activity. The Dual-Luciferase® Reporter (DLR™) Assay System (Promega Corporation) was used.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.