Luciferase assays were performed as described previously.21 (link) Briefly, cells grown in 6-well plates were transfected with an NF-κB firefly luciferase reporter plasmid 3X-κB-Luc (a gift from Dr. Hiroyuki Inuzuka) and a firefly luciferase reporter containing the IL-6 promoter (a gift of Dr. Hiroyuki Inuzuka) along with a pRL-CMV plasmid (Promega, Madison, WI, USA). After 24 h, cells were split into 96-well plates. After stimulation with aldosterone, firefly luciferase activity was measured using the Dual Luciferase Assay System and normalized to renilla luciferase activity according to manufacturer's instructions (Promega, Madison, WI, USA).
Protocol detail
NF-κB and IL-6 Promoter Luciferase Assays
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Other organizations : Fudan University, Huashan Hospital
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Variable analysis
independent variables
- Aldosterone stimulation
- Firefly luciferase activity
- Cells grown in 6-well plates
- Transfection with NF-κB firefly luciferase reporter plasmid 3X-κB-Luc
- Transfection with firefly luciferase reporter containing the IL-6 promoter
- Transfection with pRL-CMV plasmid (Promega, Madison, WI, USA)
- Dual Luciferase Assay System (Promega, Madison, WI, USA)
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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