There are robust techniques for the analysis of apoptotic and necrotic cell death modes. However, what methodology can be used to specify autophagic cell death and to discriminate this particular cell death mode from other forms?
Monica
This article is addressing ways to monitor autophagy. Let me rephrase! If you have a population of dead cells. How can you make sure that they actually died by autophagy? You can of course block the process and analyze survival but I believe most ways to inhibit autophagy are toxic to cells, at least in the long run.
Regards
Magnus
hi magnus
You can use lysosomal inhibitor such as bafilomycin A1 to block the fusion between autophagosome and lysosome.
Regards,
Luan
Dear Olsson,
Greetings! To detect autophagy you can simply use double immuno-fluorescence technique by using two anibodies LAMP and LC3B... LAMP is for lysosome and LC3B for Autophagy and then merged both image in same area. u can definitely find the autophagy. In the following article it is used to detect autophagy
http://www.tandfonline.com/doi/abs/10.1080/02772248.2013.786941#.Uzaoz6iSwfQ
I agree with Paresh. You'll have to go by the exclusion method using ZVAD for inhibition of apoptosis and necrostatin to discount necroptosis while following the LC3B and alike. Not a perfect solution, but there is none I am aware of... Good luck! You might need plenty of it.
I agree with Paresh too. I would look for inhibition of cell death with knockdown of Atg proteins. Although pharmacological inhibition of autophagy can be used (with chloroquine, bafilomycin A1 or 3MA) to look for inhibition of cell death too, these results need to be verified with a genetic manipulation of autophagy (siRNAs or shRNAs).
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