In my work am trying to construction EMT( Epithelial to mesenchymal transition) pathway induced by TGF-β but I am finding difficulties while construction i.e.,
whether pathway constructed for particular cancer is effective or in general
(because the TGF-β downstream signals are same i e SMAD and non-SMAD signal are same for all type of cancer)
what do you mean by "constructing" TGF-b induced EMT pathway? If you are looking for how to show the TGF-b induced EMT, you can show it by epithelial phenotype of cells (incubate with TGFb for 72 hours in serum free media) etc. If you need to show by western blots.....72 hours TGF-b treated cells can be lysed and probed with E-cadherin and N-cadherin antibodies. Alternatively, you could use E-cad or N-cad promoter driven luciferases or fluorescent proteins.
If you are trying to do a mathematical modeling...that is a different ball game. But some models still do exists for that.
thank you Shyam Nyati for your suggestions construction means Construction of the protein interaction network. I am trying to construct protein interaction network that induced by TGF-b during the event EMT.
Hi. You can search for your protein network (TGF-beta) with CYTOSCAPE. You could copy paste the list of your interacting proteins and then use GO ontology to look for the genes implicated in EMT process. Hope this may help you.
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