Description(s):

NFkB Responsive Luciferase Reporter NIH/3T3 Stable Cell Line is derived from mouse fibroblast,and stably express firefly luciferase reporter gene under the control of the NFkB  Response element.This cell line is an ideal cellular model for monitoring the activation of NFkB Receptor Signaling Pathway triggered by stimuli treatment, enforced gene expression and gene knockdown.

Principle

NFkB plays an important role in controlling many biological processes including immune and inflammatory responses, developmental processes, cellular growth, and apoptosis. In response to the various stimuli, such as stress, cytokines, free radicals, ultraviolet irradiation, and bacterial or viral antigens, NFkB is activated and translocates from cytoplasm to nucleus, where NFkB binds to its response element on the promoter region and regulates a wide spectrum of gene expression. Dysfunction of NFkB activity is associated with cancer, inflammatory and autoimmune disease, and viral infection. Monitoring the NFkB activity is essential to unveil the mechanism of these diseases and conduct drug discovery. Signosis has established NFkB luciferase reporter NIH/3T3 stable cell line that has been stably transfected with pTA-NFkB-luciferase reporter vector and can be used as a reporter system for monitoring the activation of NFkB triggered by stimuli treatment such as TNFα, IL-1α and IL-1β, enforced gene expression and gene knockdown. 

The cell line was established by transfection using a pTA-NFkB-luciferase reporter vector, which contains 4 repeats of NFkB binding sites, a minimal promoter upstream of the firefly luciferase coding region, along with hygromycin expression vector followed by hygromycin selection. The hygromycin resistant clones were subsequently screened for TNFa-induced luciferase activity.

 

Principle behind TF luciferase reporter.  TF luciferase reporter stable cell line utilizes artificial promoter constructs to drive luciferase expression.  The promoter region can consists of multiple repeats of a cis-element TF binding site, a DNA fragment from the promoter region of a known TF downstream gene, or a DNA fragment containing putative/known TF binding sites.  There are several ways that a TF can be activated, such as through extracellular stimuli or through intracellular signaling pathways.  Once activated, the TF translocates to the nucleus and often interacts with relevant co-factors to drive gene expression.  Once luciferase is expressed, it can generate light in an enzymatic assay and the amount of light measured is positively correlated with the level of TF activation.

 

Data

 

 

 

 

 

 

 

 

 

 

 

Analysis of SL-0006 NFκB reporter activity in response to various treatments. The cells were seeded on a 96-well plate for overnight with DMEM including 10% FBS. The cells then were treated with the following chemicals in DMEM and 0.1% FBS for 16 hours: 10ng/mL IL-1a, , 10ng/ml PMA, 10ng/ml oncostatin M (OSM), 100ng/ml IFN-gamma, and 2ng/ml TNFα.  NIH/3T3-NFkB Luciferase Reporter Cell Line exhibits response to TNFα and IL-1a when compared to untreated cells. Data was read on a luminometer with a sensitivity of 3x10^-21 moles luciferase.

Literature

View user manual

 

 

Citations

NRF2/ARE Luciferase Reporter MCF7 Stable Cell Line (SL-0010)    Withaferin A induces Nrf2-dependent protection against liver injury: role of Keap1-independent mechanisms. DL Palliyaguru, DV Chartoumpekis, N Wakabayashi. Free Radical Biology and Medicine 2016. http://dx.doi.org/10.1016/j.freeradbiomed.2016.10.003   Occurrence and in vitro toxicity of unregulated disinfection by-products in two Saskatchewan drinking water treatment plants. Christena Leigh Watts. Handle Proxy, Universidade Da Coruña, 1 Sept. 2018, hdl.handle.net/10388/11499.   Luciferase Stable Expressing Hela Cell Line (SL-0102)   Redox-responsive, reversibly-crosslinked thiolated cationic helical polypeptides for efficient siRNA encapsulation and delivery.N Zheng, Z Song, Y Liu, R Zhang, R Zhang, C Yao. Journal of Controlled Release.Volume 205, 10 May 2015, Pages 231–239   Targeted delivery of cisplatin to tumor xenografts via the nanoparticle component of nano-diamino-tetrac. Thangirala Sudha, Dhruba J Bharali, Murat Yalcin, Noureldien HE Darwish, Melis Debreli Coskun, Kelly A Keating, Hung-Yun Lin, Paul J Davis, Shaker A Mousa. February 2017 ,Vol. 12, No. 3, Pages 195-205 , DOI 10.2217/nnm-2016-0315.   NFkB Luciferase Reporter Stable Cell Lines  (SL-0001)   Thymoquinone Modulates Blood Coagulation in Vitro via Its Effects on Inflammatory and Coagulation Pathways. V Muralidharan-Chari, J Kim, A Abuawad, M Naeem.  Int. J. Mol. Sci. 2016, 17(4), 474; doi:10.3390/ijms17040474   Drug repurposing screen identifies lestaurtinib amplifies the ability of the poly (ADP-ribose) polymerase 1 inhibitor AG14361 to kill breast cancer associated gene-1 …G Vazquez-Ortiz, C Chisholm, X Xu, TJ Lahusen, C Li… - Breast Cancer Research  2014,  Jun 24;16(3):R67. doi: 10.1186/bcr3682.   NFkB Luciferase Reporter A549 Stable Cell Lines  (SL-0014)   Alcohol promotes migration and invasion of triple-negative breast cancer cells through activation of p38 MAPK and JNK†M Zhao, EW Howard, AB Parris, Z Guo, Q Zhao, X Yang.  Molecular Carcinogenesis, 2016, DOI: 10.1002/mc.22538   HEK293 TCF/LEF Luciferase Reporter Stable Cell Line (SL-0015)   Active β-catenin is regulated by the PTEN/PI3 kinase pathway: a role for protein phosphatase PP2A. Amit Persad,1,* Geetha Venkateswaran,1,* Li Hao,1 Maria E. Garcia,1 Jenny Yoon,1 Jaskiran Sidhu,1 and Sujata Persad1. Genes Cancer. 2016 Nov; 7(11-12): 368–382., doi:  10.18632/genesandcancer.128   Diallyl trisulfide inhibits proliferation, invasion and angiogenesis of glioma cells by inactivating Wnt/β-catenin signaling. Qingxia Tao, Cuiying Wu, Ruxiang Xu, Lijun Niu, Jiazhen Qin, Ning Liu, Peng Zhang, Chong Wang. Cell and Tissue Research, 2017, doi:10.1007/s00441-017-2678-9.   BCN057 induces intestinal stem cell repair and mitigates radiation-induced intestinal injury. Payel Bhanja, Andrew Norris, Pooja Gupta-Saraf, Andrew Hoover and Subhrajit Saha. Stem Cell Research & Therapy, vol. 9, no. 1, Feb. 2018, doi:10.1186/s13287-017-0763-3.   (Nrf2/ARE) luciferase reporter cell line HEK293 (SL-0042)   Direct Antioxidant Properties of Methotrexate: Inhibition of Malondialdehyde-Acetaldehyde-Protein Adduct Formation and Superoxide Scavenging. Matthew C. Zimmermana, Dahn L. Clemensb, c, Michael J. Duryeeb, Cleofes Sarmientoa, Andrew Chioub, Carlos D. Hunterb, Jun Tiana, Lynell W. Klassenb, c, James R. O’Dellb, c, Geoffrey M. Thieleb, c, Redox Biology,2017.07.018, doi: 10.1016/j.redox.2017.07.018   Active β-catenin is regulated by the PTEN/PI3 kinase pathway: a role for protein phosphatase PP2A. Amit Persad1,*, Geetha Venkateswaran1,*, Li Hao1, Maria E. Garcia1, Jenny Yoon1, Jaskiran Sidhu1 and Sujata Persad1.Genes & Cancer. January 30, 2017 . https://doi.org/10.18632/genesandcancer.128    HIF Luciferase Reporter Hela Stable Cell Line (SL-0023)   Hypothermia augments stress response in mammalian cells. Eskla, Kattri-Liis, et al. Free Radical Biology and Medicine, 2018, doi:10.1016/j.freeradbiomed.2018.04.571.   NFkB Luciferase Reporter HepG2 Stable Cell Line (SL-0017)   Hypothermia augments stress response in mammalian cells. Eskla, Kattri-Liis, et al. Free Radical Biology and Medicine, 2018, doi:10.1016/j.freeradbiomed.2018.04.571.   CHOP Luciferase Reporter Mia_Paca2 Stable Cell Line (SL-0025)   A new ent-norabietant diterpenoid from roots of Euphorbia lathyris L. Yun, Young Sook, et al.  Tetrahedron Letters, 2018, doi:10.1016/j.tetlet.2018.06.017.   NFAT Luciferase Reporter Jurkat Stable Cell Line (SL-0032)   Diversification of Bw4 Specificity and Recognition of a Nonclassical MHC Class I Molecule Implicated in Maternal–Fetal Tolerance by Killer Cell Ig-like Receptors of the Rhesus Macaque. Banerjee, Priyankana, et al. The Journal of Immunology, 2018, doi:10.4049/jimmunol.1800494.   P329G-CAR-J: a Novel Jurkat-NFAT-Based CAR-T Reporter System Recognizing the P329G Fc Mutation. Darowski, Diana, et al. Protein Engineering, Design and Selection, 2019, doi:10.1093/protein/gzz027.   Dendritic cells dictate responses to PD-L1 blockade cancer immunotherapy. Mayoux, Maud, et al.  Science Translational Medicine Vol. 12, Issue 534, 2020. DOI: 10.1126/scitranslmed.aav7431    NRF2/ARE Luciferase Reporter HepG2 Stable Cell Line (SL-0046)   Innovative drinking water treatment techniques reduce the disinfection-induced oxidative stress and genotoxic activity. Lundqvist, Johan, et al. Water Research, vol. 155, 2019, pp. 182–192., doi:10.1016/j.watres.2019.02.052.   Stat3 Luciferase Reporter Hela Stable Cell Line (SL-0003)   Discovery of a New STAT3 Inhibitor Acting on the Linker Domain. Koseki, Tatsuya, et al. Biological and Pharmaceutical Bulletin, vol. 42, no. 5, 2019, pp. 792–800., doi:10.1248/bpb.b18-00992.   An engineered ligand trap inhibits leukemia inhibitory factor as pancreatic cancer treatment strategy. Hunter, S.A., McIntosh, B.J., Shi, Y. et al. Commun Biol 4, 452 (2021). https://doi.org/10.1038/s42003-021-01928-2   Stat1 Luciferase Reporter Hela Stable Cell Line (SL-0004)   Discovery of a New STAT3 Inhibitor Acting on the Linker Domain. Koseki, Tatsuya, et al. Biological and Pharmaceutical Bulletin, vol. 42, no. 5, 2019, pp. 792–800., doi:10.1248/bpb.b18-00992.   NRF2/ARE Luciferase Reporter NIH3T3 Stable Cell Line (SL-0047)   Thiamine and Benfotiamine Protect Neuroblastoma Cells against Paraquat and β-Amyloid Toxicity by a Coenzyme-Independent Mechanism. Sambon, Margaux, et al. Heliyon, vol. 5, no. 5, 2019, doi:10.1016/j.heliyon.2019.e01710.   NFkB Luciferase Reporter HEK293 Stable Cell Line (SL-0012)   Vitale LA, He LZ, Thomas LJ, et al. Development of CDX-527: a bispecific antibody combining PD-1 blockade and CD27 costimulation for cancer immunotherapy. Cancer Immunology, Immunotherapy : CII. 2020 May. DOI: 10.1007/s00262-020-02610-y.   Luciferase Substrate (LUC15/100)   Direct Antioxidant Properties of Methotrexate: Inhibition of Malondialdehyde-Acetaldehyde-Protein Adduct Formation and Superoxide Scavenging. Matthew C. Zimmermana, Dahn L. Clemensb, c, Michael J. Duryeeb, Cleofes Sarmientoa, Andrew Chioub, Carlos D. Hunterb, Jun Tiana, Lynell W. Klassenb, c, James R. O’Dellb, c, Geoffrey M. Thieleb, c, Redox Biology,2017.07.018, doi: 10.1016/j.redox.2017.07.018   FXR Luciferase Reporter HepG2 Stable Cell Line (SL-0055) Targeting the Liver‐Brain Axis with Hop‐Derived Flavonoids Improves Lipid Metabolism and Cognitive Performance in Mice. Paraiso, I.L., Revel, J.S., Choi, J., Miranda, C.L., Lak, P., Kioussi, C., Bobe, G., Gombart, A.F., Raber, J., Maier, C.S. and Stevens, J.F. Mol. Nutr. Food Res. 2020, 2000341. https://doi.org/10.1002/mnfr.202000341   Estrogen Receptor Luciferase Reporter T47D Stable Cell Line (SL-0002)   Glass-bottled drinking water: a time capsule to study the historic presence of hazardous chemicals using effect-based methods. Lundqvist, J., Persson, K.M. & Oskarsson, A. Environ Sci Eur 33, 34 (2021). https://doi.org/10.1186/s12302-021-00476-0