JNK / AP1 Pathway
About JNK /AP1 pathway:
The stress-activated protein kinase (SAPK), Jun N-terminal kinase (JNK) family (including MAP Kinases) is activated by variety of environmental stresses. This pathway is responsible for the phosphorylation and activation of Jun. During the activation, the MAPKs activate/phosphorylate JNKs which in turn activate transcription factors in nucleus. The activated or phosphorylated transcription factors (such as c-Jun JunB and JunD) forms a protein complex with Fos protein family (c-Fos, Fos B, Fra-1 and Fra-2). The complex (the activator protein-1, AP1) binds to its response element sequence and induces the target gene transcription.
Detect JNK /AP1 pathway:
GenTarget developed a set of reporting lentivirus to monitor JNK/AP1 signaling pathway. Those reporting lentivirus has a luminescent report (Luciferase, Renilla Luc) or a fluorescent report (GFP, RFP) or a secreted SEAP report, under a minimal CMV promoter (mCMV) that embedded with optimized tandem repeats of AP1 Responsive Element (AP1-RE) sequence motif (5′- TGAGTCAG). When transcription factors bind to AP1 responsive sequence (the transcriptional response element), the downstream reporter is expressed as the result of activation for the minimal promoter.
The luciferase signal can be measured via Luciferase assay. The fluorescent reporter can be detected via its fluorescent signal. The SEAP secreted into cell culture supernatant, which allows to determine reporter activity without disturbing the cells, does not require the preparation of cell lysates and can be used for kinetic studies.
Use Report Lentivirus for JNK /AP1 pathway:
Those reporting lentivirus also constitutively expressed a fluorescent selection marker or an antibiotic selection marker under the RSV promoter, which makes it easier to select the stably infected signal reporting cells (to generate pathway specific sensor cell lines), or provides internal reference for virus transduction efficiency (when a fluorescent marker is under the RSV promoter). See the scheme below for lentivector’s core expression cassette.
The premade, ready-to-use reporter lentivirus provides a much easier tool to monitor the activity of MAPK / JNK / AP1 signaling pathways in virtually any mammalian cell type. It also allows to generate your own reporting cell line in your desired cell type for study or screen of pathway specific gene-knockdown, over-expression, or chemical / drug/protein treatment in the cell based assay. This JNK/AP1 pathway can be tested/ simulated by Phorbol 12-Myristate 13-Acetate (PMA).
Click the Product Manual (pdf) to see products with different reporter and different selection /normalization marker.