Good morning everybody,
I am currently studying the transcriptional regulation of a gene in the liver, known to be a target of HNF4α (this gene is strongly upregulated in HNF4α liver specific KO mice) in an inflammatory context (LPS).
My hypothesis is that, during inflammation, HNF4α is phosphorylated so its DNA binding activity is decreased and the transcription of my gene is not anymore inhibited by HNF4α.
So I would like to study the phosphorylation status of HNF4α in my case but I really don’t know which precise phosphorylation I need to look (serin or threonin?, localization ?).
Do you know a good antibody to do this by Western Blot ?
Hi. Only by curiosity could it be also the opposite ? HNF4 could be dephosphorylated during inflamation ? I believe it is not the phosphate by itself which modifies the affinity for the promotor but the structural change of the transcription factor due to phosphorylation or dephosphorylation which makes that it does not bind anymore to the promotor. I suppose that using 32P it is possible to identify the phosphorylated HNF4 during inflamation.