I transfected my cells with a vector containing Wnt5a gene. This stable transfection changed my behavior of cells specifically. Due to the long-term expression of Wnt5a in my cells, Do use of Wnt5a antagonist in the short term, can antagonize cells behavior?
Hi Vajihe,
it depends on what you want to do with your experiment. If what you need is a rescue of your phenotype you might use inhibitors of Porcupine (such as Iwp2), this will inhibit secretion of all Wnts (including your overexpressed Wnt5a). You can also silence the Wnt5a receptor or using blocking antibodies if available, but once again it depends on what you ask from your experiment.
Pleas consider that Wnt overexpression affects not only transfected cells, but also untransfected surrounding cells in which Wnt5a acts trhough paracrine mechanism.
By the way the morphological phenotype is going to be reverted quite soon I suspect ;)
All the best, hope it helps
Francesco
I agree with Francesco - first use an inhibitor of Wnt secretion- IWP2 or C59 and see if the effect is lost or reduced. OR Can you get the same effect by just adding Wnt5a conditioned medium to the untransfected cells?
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