Hypoxia - downregulated HIF-1A - why me??? (Apr/15/2010 )

Hi all..

I have been tryin to study the regulation og HIF-1A for my hypoxic experiments. I tried doing cobalt chloride dose response, hypoxia incubators and bubbling up culture media with Nitrogen, and still HIF-1A is not up regulated in my cells. My other protein of interest which is a hypoxia response protein is upregulated, so to confirm that the cells are undergoing hypoxia, HIF-1A the marker for hypoxia is not upregulated...

can anybody pleeeeeeeeeeeeeeease suggest why this is the case and ways to rectify the problem??? I have been stuck with this for the past 2 years!!!! yes 2 precious years of my PhD!!!!!!!!!!!11

Many Thanks...

Hi!
A question, what do you look for? HIF mRNA or protein??? Because under hypoxia, the first thing that happens is an stabilization of HIF protein, not an induction of HIF transcription... Sorry about my English

Yes, the regulation of HIF-alpha activity is done mainly on the protein level. The expression is constitutive, but may be upregulated in cancers or long term hypoxia.