I am setting up an in vitro model using macrophages to induce pyroptosis via inflammasome activation and want to know how long this will take, and what should I be looking for?
In a short time (1-3 h) (see the link below). One can follow LDH release from cells using a kit.
Link: http://www.ncbi.nlm.nih.gov/pubmed/21884178
It will largely depend on what induction model you are using and whether you are viewing the cells by a microscope or reading out by some biochemical method (LDH release, WB for processing, IL1b, etc).
In LPS-primed J774A.1 stimulated with ATP you can see the first cells pyroptosing within 10-20 minutes. In the same cells stimulated with nigericin you can see them begin to pyroptose at about 20-30 minutes. However, you would only be able to sensitively detect this using LDH at about 45-60 minutes. It's a ramp-up curve.
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