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Immunofluorescent Staining for ASC Speck Formation
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To visualize ASC speck formation in response to different treatments, cells cultured on 24-chamber slides were fixed with 4% PFA for 10 min and then permeabilized with 0.3% Triton X-100 for 15 min. Cells were then blocked with 3% donkey serum and stained with anti-ASC (AdipoGen, AG-25B-0006) at 4 °C overnight. Then, Alexa-Fluor 555 conjugated secondary antibody was used. The slides were monitored using a fluorescence microscope (TS SPE; Leica Microsystems, Germany).
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