HEK293T cells were cotransfected with a firefly luciferase reporter construct under the control of three NF-κB binding sites55 (link), a Gaussia luciferase construct55 (link) for normalization, and expression vectors for a constitutively active mutant of IKKβ56 (link) and the functional vpu alleles as described15 (link). Two days post-transfection, Gaussia luciferase activity in the supernatant was measured using the Gaussia-Juice Kit (p.j.k.) and the cells were used to measure firefly luciferase activity using the Luciferase Assay from Promega with an Orion Microplate luminometer (Berthold).
Other organizations :
Universität Ulm, University of Pennsylvania, German Primate Center, Innsbruck Medical University, Emory University, University of Pittsburgh, Institut Pasteur
Firefly luciferase reporter construct under the control of three NF-κB binding sites
Gaussia luciferase construct for normalization
controls
Positive control: Constitutively active mutant of IKKβ
Negative control: Not explicitly mentioned
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