You can add LPS 100-250 nM for 16 hrs followed with 5 mM ATP or Nigericin 40 micromolar for 2-5 hrs (need to check the cells every hour) to Raw cells/BMDM.

Thanks

Thanks, Debanjan Mukhopadhyay, for the help.

Your method worked well for identifying swelling cells after Nigericin treatment. These cells underwent membrane rupture after higher dose treatment. However, it seems that only about 10% of the cells have an ASC speck in the perinuclear region, while others have a smear ASC signal in the nucleus and cytosol. I think there still might be something wrong with my assay.