I am working to understand how SHH pathway is involved in endothelial differentiation from human embryonic stem cells. I use both recombinant SHH and SANT1 to activate and inhibit the pathway.I find that by qRT PCR SMO expression does not change, but PATCHED1 expression changes depending on whether the pathway is active or inactive.
In you expert opinion, can PATCHED1 expression be used to understand if recombinant SHH has activated the pathway or SANT1 has inhibited the pathway?
Also in your opinion, which is the better molecular biology technique to study the sonic hedgehog pathway?
I am eagerly waiting for your answer.
Thanks.
Regards,
Prasad.
Hi Prasad,
I am not sure if Patched1 alone expression would be enough for indicating the Ssh pathway activation or inhibition. Usually, people showed phosphorylation of downstream kinases. In that respect, using a kinase activity profiling of your samples would be much more spot-on proof. For the same, I would recommend the unique functional-based kinome activity profiling from PamGene ( https://pamgene.com/science/) which covers all the kinases involved in Ssh pathway activation/inhibition for complete pathway elucidation.
All the best,
Tushar
Patch1 generally is upregulated in response to activating shh signaling. It has been used as a marker for activation in many of the older papers.
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