Hi all,
I'm currently looking at the dynamics of P-bodies in human cultured cell lines/tissues by immunofluorescence. As a comparison to my positive cell lines, I was interested to use (if available) a cell line that does NOT exhibit P-bodies, and/or is naturally unable to assemble them.
I've performed a search of the literature (white and grey) and haven't been able to find any report of a cell line like this. Could anyone out there suggest to me any cell line(s) and/or body tissue(s) that do NOT form P-bodies, and if so what conditions they need to be cultured under?
Thanks!
Dan
(Dr. Daniel Scott)
Hey Dan!
The "endogenous control" you're suggesting would likely be the most elegant approach but may be tough to find.
As an alternative, you could think of disrupting the assembly of P-bodies through more than a single "exogenous" approach.
Check out the link to an RNA biology paper describing a screen for myxobacterial compounds which inhibit P-body assembly. Gephyronic acid, a protein synthesis inhibitor, efficiently blocks P-body assembly. In addition to a pharmacological approach, you could envision a DDX6 siRNA.
These controls may be weaker than a cell line naturally unable to assemble P-bodies, if it exists, but providing two different perspectives may make the conclusions somewhat stronger.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3907476/
Hi Fabio!
Thanks for your suggestions - they're very helpful! I will definitely have a siRNA-mediated control as you suggest, but was just hoping that there might be a natural (biological) control that I could add as well.
The paper that you include looks really interesting and useful - thanks for suggesting it!
Cheers,
Dan
Cool! Your new ALS project sounds compelling. I'm excited to hear about it at some Axis or Interlab meeting ;)
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