Quantifying Cellular Signaling Pathways

The DLR assays were performed as described previously (38 (link), 40 (link)–42 (link), 53 (link)). In short, HEK293T cells were plated on 24 well dish (Corning) at a density of 1 × 10⁵ cells per well-overnight before transfection. Cells were then co-transfected with 100 ng of the indicated expression plasmid, 100 ng of IFN-β or NF-κB promoter reporter and 10 ng of pRL-TK (internal control) to normalize transfection efficiency. Twenty-four hours post-transfection, the luciferase activity was detected with a luciferase assay kit (Promega).

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Li M., Liao Z., Xu Z., Zou X., Wang Y., Peng H., Li Y., Ou X., Deng Y., Guo Y., Gan W., Peng T., Chen D, & Cai M. (2019). The Interaction Mechanism Between Herpes Simplex Virus 1 Glycoprotein D and Host Antiviral Protein Viperin. Frontiers in Immunology, 10, 2810.

Publication 2019

24 well dish luciferase assay kit

Assay Cells Dish Luciferase Nf κb Plasmid Promega Transfection

Corresponding Organization : Second Affiliated Hospital of Guangzhou Medical University

Other organizations : Yue Bei People's Hospital, Shantou University, State Key Laboratory of Respiratory Disease

Top 5 similar protocols

Protocol cited in 2 other protocols

Variable analysis

independent variables

Expression plasmid

dependent variables

Luciferase activity

control variables

HEK293T cells
24-well dish (Corning)
Cell density (1 × 10^5 cells per well)
Transfection efficiency (normalized by pRL-TK)

positive controls

IFN-β promoter reporter
NF-κB promoter reporter

negative controls

Not explicitly mentioned

Annotations

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