The 3x-Bcl6 reporter construct has been described previously9 (link). Slc2a3, Slc2a1, P4ha2, Plod2 and Tpi1 reporter constructs were prepared by cloning the promoters of each gene into the pGL3-basic luciferase reporter construct (Promega). EL4 or A20 cells were co-transfected with the indicated expression vectors along with the promoter-reporter constructs as well as a CMV-renilla control plasmid to normalize for transfection efficiency. The samples were harvested 16–24 h post-transfection and then analyzed with the Dual-Luciferase Reporter system (Promega).
Protocol detail
Gene Promoter Luciferase Assays
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Corresponding Organization : University of Washington
Other organizations : Carilion Clinic, Virginia Tech, University of Alabama at Birmingham
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Variable analysis
independent variables
- Expression vectors
- Luciferase reporter activity
- CMV-renilla control plasmid to normalize for transfection efficiency
- Positive control: 3x-Bcl6 reporter construct
- Negative control: Not explicitly mentioned
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