Luciferase reporter gene vector pGL4 containing the PGC-1α promoter area was obtained from Baihao Bio (China, Shenyang). pGL4-PGC-1α or control was transfected into THP-1-derived macrophages. Reporter assays were conducted 24 h after administrating with catalpol (0, 5, 20, and 80 μM) or pioglitazone (5 μM). The luciferase activity was determined with a Dual-Luciferase Reporter Assay Kit (Promega, Wisconsin, USA) using a Dual-Light Chemiluminescent Reporter Gene Assay System and was normalized to the Renilla luciferase activity.
Dual light chemiluminescent reporter gene assay system
Manufactured by Promega
Sourced in United States
The Dual-Light Chemiluminescent Reporter Gene Assay System is a laboratory equipment product that measures the activity of two reporter genes simultaneously. It utilizes chemiluminescent technology to quantify gene expression levels.
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Lab products found in correlation
2 protocols using dual light chemiluminescent reporter gene assay system
1
Catalpol Modulates PGC-1α Promoter Activity
2
Validating the miR-23a-3p-MAP4K4 Interaction
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In order to verify the relationship between miR-23a-3p and MAP4K4, we inserted the 3'UTR region of MAP4K4 into a vector and transfected the vector into 293T cells. Additionally, we performed site-directed mutagenesis to further determine the site of action of the miRNA and the 3' UTR of MAP4K4. Luciferase activity was determined with a Dual-Luciferase Reporter Assay Kit using a Dual-Light Chemiluminescent Reporter Gene Assay System (Promega, Fitchburg, WI) and was normalized to the Renilla (ObiO Co., Ltd, Shanghai, China) luciferase. The rat MAP4K4 3'UTR was ampli ed and cloned into a pGL4 vector containing the re y luciferase reporter gene (ObiO Co., Ltd, Shanghai, China). For the luciferase assay, 293T cells were cotransfected with 100ng re y luciferase constructs, 10 ng pRL-TK Renilla luciferase plasmid, and 100 nmol/L synthetic miR-23a-3p mimic and no-load vector. The results are expressed as relative luciferase activity (Fire y luciferase/Renilla luciferase).
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