Leading Lightâ Wnt Reporter Cell Line

S4b c producing a range of reporter activity similar.

Leading lightâ wnt reporter cell line.

Wnt proteins bind to receptors on the cell surface initiating a signaling cascade that leads to stabilization and nuclear translocation of beta catenin. The wnt pathway controls a large and diverse set of cell fate decisions in embryonic development adult organ maintenance and disease. The wnt signaling pathway helps animal cells to communicate with each other to coordinate the formation of tissues and organs. The leading light wnt reporter cell line is the basis for a cell based luciferase activity test suitable for a multi well plate format such as 96 or 384 well microplates.

The system contains an engineered 3t3 mouse fibroblast cell line which expresses the firefly luciferase reporter gene under the control of wnt responsive promoters tcf lef. We found that both colon cancer cell lines induced canonical wnt signalling in a cell number dependent manner fig. Beta catenin then binds to tcf lef transcription factors in the nucleus leading to transcription and. To enhance the reporter responsiveness these regulatory dna elements are synthesized in tandem repeats.

The active wnt reporter cell line is designed to monitor the activity of beta catenin based wnt signal transduction pathway. The pathway relies on a protein called wnt that is released from cells and binds to a receptor protein called frizzled on the surface of other cells to trigger changes in gene activation. There are various reporter transgenes that respond to wnt signals in intact animals and therefore as far as one can know reflect endogenous wnt signaling reviewed in barolo 2006 these reporters are based on a multimerized tcf binding site driving expression of lacz or gfp. Since the tcf lef h2b gfp transgenic is the first wnt reporter line to reveal the dynamics of wnt signaling activity in cells of the visceral endoderm of e5 5 embryos we focused on this stage to study the behavior of cells expressing the reporter.

The leading light wnt cell line assay reagents are intended for use with the leading light wnt reporter cell line which is the basis for a cell based luciferase activity test enz 61001 leading light wnt reporter assay starter kit suitable for a 96 well plate format. To monitor wnt signaling in intact cells human embryonic kidney hek293 cells were transiently transfected with a tcf lef luciferase reporter plasmid figure 1a as basal wnt signaling is relatively low in these cells figure s1a the gsk3 inhibitor chir99021 was used to enhance β catenin stabilization and activate wnt signaling the tankyrase inhibitor xav939 a known wnt signaling. The system contains an engineered 3t3 mouse fibroblast cell line which expresses the firefly luciferase reporter gene. This is for instance the case of the first zebrafish wnt reporter line which was created through the juxtaposition of four dna regulatory elements specifically binding the tcf lef complex in wnt responsive cells dorsky et al.